Adsorbens and devices for the reduction of small organic compounds from blood products

TECHNICAL FIELD

0001the This invention relates to elements for the reduction of small organic compounds in blood products.

BACKGROUND

0002On extensive research with respect to the volume of removal of substances from blood products is available. The bulk of this research is directed to the reduction of white blood cells. See, eg. BMN . Boomgaard et al, Transfusion 34: 311 (1994; F. Bertolini et al. Vox Sang 62: 82 (1992); and AM-Joustra Dijkhuis et al, Vox Sang. 67: 22 (1994). Filtration of platelets is the most commonly method used to reduce the white blood cells in platelet concentrates is. See, eg. BM Böck Et al., Transfusion 31: 333 (1991) (SEPACELL PL-5A, Asahi, Tokyo, Japan); . JD Sweeney et al, Transfusion 35: 131 (1995) (Leukotrap PL, Miles Inc., Covina, CA) and M. van Marwijk et al., Transfusion 30: 34 (1990) (CellSelect, NPBI, Emmer-Compascuum, The Netherlands; Immugard Ig 500, Terumo, Tokyo, Japan). This current filtration mechanisms but not for removing relatively low molecular weight compounds usable including for example psoralens Photoaddition products and other compounds, often occur in the treatment of biological fluids.

0003Of the Adsorption was used to isolate selective blood components used to Phospholipidpolymeren. For example, various Copolymers with different electrical charges on their interactions evaluated with blood components, including platelet adhesion and Protein adsorption. K. Ishihara et al., J. Biomed. Mat. Res. 28: 1347 (1994). However, such polymers are not on the adsorption low molecular weight compounds designed.

0004Various Dialysis methods are capable of low molecular weight compounds to remove from plasma and whole blood. For example, the dialysis low molecular weight toxins and pharmaceutical compounds successfully remove. Thus could dialysis to remove, for example, psoralens and Psoralenphotoprodukten be used from blood products. Unfortunately, reflect the current Dialysis process very complicated and expensive elements a. As such would be the Use of dialysis machines for the decontamination of large quantities of blood products not practical.

0005the Using Polystyrenedivinylbenzene, silica gel and acrylic ester polymers for the Adsorption of methylene blue was already described before. To the Example describes PCT Publication Nos. WO 91/03933 batch tests with Wi-adsorption (Z. B. Amber Lites (Rohm and Haas (Frankfurt, Germany) and Bio Beads (Bio-Rad Laboratories (Munich, Germany)). Without very careful removal of Adsorbenzharze, after they were exposed to the blood product containing these However, methods the risk of transfusion of resin particles.

0006In addition, were also elements and processes for removing leukocytes and viral inactivating agents (eg. as psoralens, hypericin and dyes such as methylene blue, toluidine blue and crystal violet) described. Specifically, in PCT Publication No. WO. 95/18665 a filter described comprising a laid textile fabric, which contains a mechanically stable polymeric substrate. The Fabric itself comprises winding textile fibers, the matrix a with interstices form and fibrillated particles that are incorporated in the interstices are. However, this element causes a significant decrease in Factor XI activity.

0007In WO 96/39818 is the use of a compound having a nucleic acid binding Ligand and a mustard group to inactivate pathogens in describes a biological material. may After treatment the removal of the chemical product using a adsorbent be made.

0008Therefore are simpler, safer and more economical methods of reducing the concentration of small organic compounds in a biological liquid while substantially maintaining the biological activity of the purified liquid required.

DESCRIPTION OF THE INVENTION

0009the present invention provides an adsorbent medium for reducing the concentration of a small organic compound in a Blood product while substantially maintaining a desired biological activity the blood product comprising (1) highly porous adsorbent that for Adsorbing the small organic compound capable of and (2) a sintered polymeric matrix, wherein the adsorbent particles are immobilized.

0010the Immobilization of adsorbent in an inert matrix for Reducing the concentration of small organic compounds in Blood products has several benefits and advantages over the existing systems. One advantage is the reduced leakage of loose adsorbent particles in the blood products, which in turn a produces safer blood products. Another advantage is the increased adsorption capacity of the Adsorbent of small organic compounds, partially due to the elimination of Verklumpungsproblemen, the immobilized non-with Particles for use with blood products, eg. As materials Red blood cells are include, connected. A further advantage is the reduction problems associated with handling loose adsorbent particles, whereby the production process is simplified. A surprising Benefit through the use of immobilized on an inert matrix adsorbent caused by a reduced damage of stored membrane containing Blood products, for example platelets, red blood cells and white Blood cells.

0011In one embodiment , the adsorbent has a surface area of ​​greater than about 750 m²/ G. In another embodiment, can the adsorbent particles have a surface area of ​​greater than about 1000 m²/ G possess.

0012In one embodiment is the element a flow element, wherein the adsorbent have a diameter of less than about 200 microns and larger than 10 microns. In an alternative embodiment is the element a charge element, wherein the adsorbent have a diameter of less than 1200 microns and larger than 300 microns.

0013at of the present invention, it is contemplated that the adsorbent polyaromatic compounds or activated carbon include. In a embodiment the polyaromatic compounds in hyper crosslinked polystyrene divinylbenzene copolymer networks contain. In one embodiment, the activated carbon may be derived from a natural source. In Alternatively, the activated carbon may be from a synthetic source from treatment of highly sulfonated polystyrene beads by means of a high-temperature activation process result.

0014furthermore is considered an element which concentration the of acridines, acridine derivatives, Methylenbau or thiols in a Blood product reduces. is also contemplated an element which for the concentration of psoralens, psoralen derivatives, including Example 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, Isopsoralenen or aminopsoralens reduced. The blood product can chosen be from a group consisting of platelets, red blood cells or plasma.

0015As an additional Component is pulled in the invention contemplated that the member also includes a blood bag.

0016The Adsorption medium and the elements of the invention may be used a method of reducing the concentration of cyclic compounds in a blood product while substantially maintaining a desired biological activity the blood product is used which method the following Steps: a) contacting the blood product with the elements of the present invention, and b) removing the blood product from the contact with the elements of the present invention.

0017In addition, an element taken into consideration that the concentration of acridines Acridine derivatives, methylene blue of thiols in a blood product reduced. is also contemplated an element which the concentration of psoralens, psoralen derivatives, including, for Example 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, Isopsoralenen or aminopsoralens reduced. The blood product can chosen be selected from the group consisting of platelets, red blood cells or plasma.

0018As an additional Component is pulled in the invention contemplated that the member also includes a blood bag.

0019Also describes a method for inactivating pathogens in Solution, which method comprises: a) providing, in any order: i) a cyclic compound, ii) a solution suspected of having the contaminated pathogens, and iii) a fibrous resin; b) treating the solution with the cyclic compound to a treated product solution to obtain, wherein the pathogens are inactivated; and c) contacting the treated product solution with the fibrous Resin, wherein the improvement is an element for reducing the concentration of small organic compounds in a blood product while substantially Maintaining a desired biological activity includes the blood product, the element highly porous adsorbent and wherein the adsorbent particles by an inert matrix are immobilized.

0020Also A method is described for reducing the concentration of cyclic compounds in solution, comprising: a) providing i) cyclic compounds in solution at a concentration, wherein the cyclic compounds are selected from the group consisting of monocyclic compounds, polycyclic Compounds and heterocyclic compounds, and ii) a fibrous resin; and b) contacting the cyclic compounds with the fibrous resin, thereby reducing the concentration of cyclic compounds.

0021Also described is a blood bag comprising; a) a biocompatible housing; and b) in a fiber network immobilized resin, the immobilized Resin is received within the biocompatible housing.

BRIEF DESCRIPTION OF THE DRAWINGS

0022In <figref idrefs="S117">1</figref> is a perspective view of an embodiment of a fiber as Graph, showing their inner core and the outer sleeve, which the fiber network of the fibrous Resin forms.

0023In <figref idrefs="S118">2</figref> is a portion of an embodiment of the fibrous Resin of the present invention is schematically shown.

0024In <figref idrefs="S119">3</figref> is a cross-sectional view of an embodiment of the fibrous resin shown in the diagram, wherein the Adsorbenskügelchen anchored to fibers are that the fibrous forming resin.

0025In <figref idrefs="S120">4</figref> is a cross-sectional view of an embodiment of the fibrous resin shown in the diagram, wherein the fibers within the Adsorbenskügelchen of the fibrous Resin are immobilized, as well as the heat seals, the samples of the fibrous surrounded resin.

0026In <figref idrefs="S121">5</figref> is is a graph showing a comparison of adsorption kinetics for removing aminopsoralens of platelets with loose Adsorbenskügelchen of Dowex^® XUS-43493 and Amberlite^® XAD-16 HP with a fibrous shown resin Amberlite^® XAD-16 contains.

0027In <figref idrefs="S122">6</figref> is is a graph showing a comparison of adsorption kinetics for removing aminopsoralens of platelets with fibrous resin, the Amberlite^® XAD-16 contains, with fibrous Resin with two different loadings of activated carbon shown.

0028In <figref idrefs="S123">7</figref> is is a graph showing a comparison of adsorption kinetics -coated for removing aminopsoralens of platelets with p (HEMA) and uncoated Dowex^® XUS-43493 beads shown.

0029In <figref idrefs="S124">8th</figref> is a graphical representation of a comparison of the effect of Glycerolgehalts in a pretreatment solution to the relative adsorptivity of aminopsoralens for Amberlite^® XAD-16 and Dowex^® XUS-43493 shown.

0030In <figref idrefs="S125">9</figref> is a graphical representation of a comparison of the effect of a wetting solution to adsorptivity of 4 '- (4-amino-2-oxa) butyl-4,5', 8- trimethylpsoralen for dried Adsorbent in 100% plasma for Amberlite^® XAD-16 (Below) and Dowex^® HXUR-43493 (above) shown; the samples that were not wetted in an ethanol solution are identified with "no Tx". The adsorptivity is expressed as a percentage relative to the capacity of optimally wet adsorbent indicated.

0031In <figref idrefs="S126">10</figref> is a graphical representation of a comparison the adsorption of aminopsoralens over a period of 3 hours from plasma using Amberlite®^® XAD-16 shown that in several various solutions have been wetted.

0032In <figref idrefs="S127">11</figref> is a graphical representation of a comparison the adsorption kinetics of methylene blue over a period of 2 hours shown from plasma.

0033In <figref idrefs="S128">12</figref> are the chemical structures of acridine, Acridine orange, 9-amino acridine and 5 - [(β-Carobxyethyl) amino] acridine shown.

0034In <figref idrefs="S129">13</figref> is a graphical representation shown in the data for adenine capacity (y-axis) and 5 - [(β-carboxyethyl) amino] acridine capacity (x-axis) for different Resins are applied.

0035In <figref idrefs="S130">14A</figref> and <figref idrefs="S131">14B</figref> is is a graph showing a comparison of adsorption kinetics for Removal of 5 - [(β-carboxyethyl) amino] acridine with Dowex^® XUS-43493 and Purolite^® MN-200 and Amberlite^® XAD-16 HP shown.

0036In <figref idrefs="S132">15</figref> is a graphical representation of a comparison the adsorption kinetics for the removal of 9-amino acridine and acridine orange with Dowex®^® XUS-43493 shown.

0037In <figref idrefs="S133">16</figref> is an illustration of the flow and batch configurations the immobilized adsorption (IAD) shown.

0038In <figref idrefs="S134">17</figref> is a graphical representation of a comparison the adsorption isotherms different Ambersorbs shown in comparison to Purolite MN-200th

0039In <figref idrefs="S135">18</figref> is a graphical representation of a comparison acridine [(β-carboxyethyl) amino] - the amount of 5 and GSH in the supernatant 300 ml PRBC units with continued or after 24 hours completion suspending a fibrous Pica G277 IAD (500 g / m²) about a storage period of 4 weeks at 4 ° C is shown.

0040In <figref idrefs="S136">19</figref> is a graphical representation of the effect acridine [(β-carboxyethyl) amino] - of including material on 5 Exposed to PRBC shown in the IADs.

0041In <figref idrefs="S137">20</figref> is a graph showing a comparison the percentage hemolysis for the non-immobilized and immobilized adsorbent Purolite MN-200 is shown.

0042In <figref idrefs="S138">21</figref> is a graphical representation of a comparison the percentage hemolysis for the non-immobilized and immobilized activated carbon adsorbent Pica G-277 shown.

0043In <figref idrefs="S139">22</figref> is a graphical representation of the kinetics for the Removal of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylenpsoralen platelet concentrates shown.

BEST MODE THE INVENTION

0044the present invention provides the materials and elements to reduce the concentration of small organic compounds from a treated blood product prepared. The small organic compounds can cyclic and acyclic compounds include. Examples of the compounds include Pathogen inactivating Compounds, dyes, and thiols. It provided elements, the immobilized a three-dimensional network of adsorbent, by an inert matrix include. By this immobilization the risk of leakage of loose adsorbent particles in the blood product reduced. also simplifies the immobilization of adsorbent in an inert Matrix production, by the loose with the handling adsorbent related problems are reduced. The immobilization of adsorbent elevated the adsorption capacity the adsorbent of small organic compounds in compositions, the red blood cells and / or platelets and / or plasma. Finally, the use of reduced of adsorbent particles immobilized by an inert matrix, injury of the compositions, the cellular membranes, z. B. platelets, Red blood cells and white blood corpuscle containing blood products include.

DEFINITIONS

0045Of the Term "acridine derivatives" refers to a chemical compound having the tricyclic structure of acridine (dibenzo [b, e] pyridine; 10-Azanthracen) contains. The compounds have an affinity for nucleic acids (and can Intercalation non-covalently bind to). The term "amino acridine" refers to acridine compounds with one or more nitrogen-containing functional groups. Examples of aminoacridines include 9-amino acridine and acridine orange (Shown in <figref idrefs="S128">12</figref>).

0046Of the Term "adsorbent" refers broadly to to any natural or synthetic material to interact with molecules in a liquid capable whereby removal of the molecule from the liquid possible becomes. Examples of naturally occurring adsorbents include, limited without it to be, activated carbon, silica, diatomaceous earth and cellulose. examples the synthetic adsorbents include, limited without it to be, polystyrene, polyacrylics and carbonaceous adsorbents. Adsorbent particles are often porous, often have large surface areas and can a plurality of functional groups (for. example, ionic, hydrophobic, acidic, basic) to be modified, which impact on can, as the adsorbent with the molecules interacts.

0047Of the Term "aromatic", "aromatic compounds", and the like refers broadly to compounds containing rings of atoms with delocalized have electrons. The monocyclic compound benzene (C₆H₆) Represents conventional aromatic compound. The Eletronen delocalization but can more as an adjacent ring occur (eg. as naphthalene (two rings) and anthracene (three rings)). At various classes of aromatic include compounds limited without it to be, aromatic halides (aryl halides), aromatic heterocyclic Compounds, aromatic hydrocarbons (arenes), and aromatic Nitro compounds (aryl nitro compounds).

0048Of the Term "biocompatible Coating "refers broadly to the cover a surface (eg. as the surface of a Polystyrene bead) with a hydrophilic polymer which when in contact with a blood product not to a harmful, toxic or immunological reaction results and the surface biocompatible makes by cell adhesion, the and reduced protein adsorption and improved cell function. Suitable coatings are biocompatible if they have a minimum, or no, effect on them to be exposed to biological show material. By "minimal" effect is meant, that no significant difference compared to control it recognizable is. In preferred embodiments, improved biocompatible coatings Oberflächenhämokompatibilität of polymeric Structures. For example, is often Poly (2-hydroxyethylmethacrylate) (pHEMA) for coating materials used in medical elements (eg., blood filters) using find.

0049Of the Term "biocompatible Housing "refers broadly to on filter housing, Container, Bags, vessels, containers and similar, which for receiving a biological material such as whole blood, Platelet concentrates and plasma are. Suitable containers are biocompatible if they have a minimal, or no, effect point to the contained therein biological material. By "minimal" effect is meant, that no significant difference between the blood product function compared to the control, as described herein, for red blood cells, platelets and plasma can be seen. Therefore, blood products may be used in biocompatible housings are mounted to a recipient prior to transfusion. In preferred Embodiments are biocompatible housing Blood bags, including a platelet storage container.

0050Of the Term "biological Liquids "includes human or non-human whole blood, plasma, platelets, red blood cells, Leukocytes, serum, lymph, saliva, milk, urine or products, derived from any of the above or are contained therein, and singly or in combination with or without a chemical additive solution. Preferably is the liquid Blood or a blood product with or without a chemical additive solution, preferably Plasma, platelets or red blood cells, most preferably apheresis plasma and red blood cells.

0051Of the Term "blood bag" refers to a blood product container.

0052Of the Term "blood product" refers to the liquid and / or associated cellular Elements and the like (Such as. For example, erythrocytes, leukocytes, platelets, etc.), the by Circulatory system of the body move; Blood products include, but are not limited to, blood cells, platelet mixtures, Serum and plasma. The term "platelet mixture" refers to one type of blood product wherein the cellular element is primarily or only platelets. A platelet concentrate (PC) is one type of platelet mixture, wherein the platelets are connected to a small reindeer than normal portion of plasma. can in blood products synthetic media supplement that volume that normally by plasma occupied; for example, a platelet concentrate may in 35% plasma / 65% synthetic media suspended platelets include. Frequently includes the synthetic medium phosphate.

0053Of the Term "blood separation device" refers broadly to an item, machine or the like, the for separating blood into blood products (eg. as platelets and Plasma) capable is. An apheresis system is one type of blood separation device represent. apheresis systems generally comprise a blood separation member, a intricate network of tubes and filtering, collection bags, an anticoagulant, and a computerized Device for controlling all of the components.

0054Of the Term "crosslinked" refers broadly to to linear molecules, which are bonded to each other and thereby a two or three dimensional forming network. For example serves divinylbenzene (DVB) as the Crosslinking agent in the formation of styrene-divinylbenzene copolymers. Of the Term also includes "hypercrosslinking" said hypercrosslinked Networks by crosslinking linear polystyrene chains either in solution or generated in a swollen state with bifunctional agents will. A variety of bifunctional agents can be used for networking be used (see, for. example, Davankov and Tsyurupa, Reactive Polymers 13: 24-42 (1990); . Tsyurupa et al, Reactive Polymers 25: 69-78 (1995).

0055Of the Term "cyclic Compounds "refers to compounds having one (ie, a monocyclic compound) or more than one (ie, polycyclic compound) ring of atoms. The term is not limited to compounds with rings a certain number of atoms. While most cyclic Compounds rings with five contain or six atoms, and rings with different numbers are of atoms (eg. for example, three or four atoms) by the present invention taken into consideration. The identity the atoms in the rings is not limited, although the atoms usually predominantly Are carbon atoms. Generally speaking, limits the rings polycyclic compounds together to; all recently includes the Term "polycyclic" compound also Compounds containing multiple, non-contiguous rings.

0056Of the Term "dye" refers broadly to on compounds which impart color. Dyes generally comprise chromophores and auxochromic groups that are cyclic, in one or more Compounds are attached. The color is due to the chromophore, whereas the Färbeaffinitäten on the auxochrome are due. Dyes have been grouped in many categories, including Azine dyes (eg neutral red, safranin and Azocarmine B.); the Azo dyes; the Azocarmine dyes; the diphenylmethane dyes, fluorescein dyes Ketonimin dyes, which dyes rosaniline and triphenylmethane dyes. It is contemplated, that the methods and elements of the present invention in conjunction can be performed with any dye which a cyclic compound.

0057Of the Term "fibrous resin" generally refers the immobilization of adsorbent material, including, for Example of resins in, on or entrapped to a fiber network. In one embodiment, made up the fiber network of polymeric fibers. At a made another embodiment the fibers of a polymer core (eg., polyethylene terephthalate [PET]) with a high melting point of a polymer sleeve (z. As nylon or modified PET) with a relatively low melting temperature surrounded. fibrous Resin may be produced by heating the fiber network under conditions be, the adsorption capacity, the the resin affect to any significant degree. Where the resin globule comprises a heating is performed so that the Adsorbenskügelchen bonded to the outer polymeric sleeve be to create "fibrous beads". By Production of fibrous Resin containing a known amount of Adsorbenskügelchen per defined area, can sample of fibrous Resin for use in the removal of cyclic compounds (Z. B. psoralens, and in particular aminopsoralens) and other Products by cutting out a defined area of fibrous Resin, instead of weighing the Adsorbenskügelchen obtained.

0058Of the Term "filter" refers broadly to to elements, materials, and the like, The particular components of a mixture permit the passage can and thereby restrain other components. For example, a filter, a mesh with pores in a Size include, which a blood product (eg., plasma) allow the passage and while other components such as resin particles restrain. The term "filter" is not limited to the Member limited, be retained by certain compounds.

0059Of the Term "flow adapter" refers to an element for controlling the flow of a particular Substance, eg. As a blood product is capable of. The Durchflussadaptor may include additional fulfill functions, z. B. the passage of pieces prevent the Adsorptionsharzmaterials.

0060Of the Term "heterocyclic Compounds "refers broadly to cyclic compounds wherein one or more the rings more than one type of atom. In general terms, Carbon is the predominant Atom, said to the other atoms for example, nitrogen, sulfur and oxygen include. To Examples of heterocyclic compounds include furan, pyrrole, thiophene and pyridine.

0061the Called "high-temperature activation process" refers to a high-temperature process, which typically changes at surface, porosity and surface chemistry of the treated material due to pyrolysis and / or oxidation the starting material leads.

0062the Label "immobilized Adsorption (IAD) "refers be included on an immobilized adsorbent material in, on or to an inert matrix. Where the inert matrix a fiber network is the term IAD can be interchangeable with the term fibrous resin be used.

0063Of the Term inert matrix refers to any synthetic or Naturally occurring fiber or polymeric material for immobilizing can be used by adsorbent particles without the desired biological activity the blood product substantially affecting. The matrix can be used to Reducing the concentration of small organic compounds contribute, although they typically are not essential to the adsorption or removal process contributes. also can interact the inert matrix with cells or protein components, leading to cell removal (z. B. leucodepletion) or removing Protein, or other molecules leads.

0064Of the Term "in-line column" refers to a container of usually cylindrical shape having an inlet end and an outlet end, contains a substance incorporated therein, the concentration of small reducing organic compounds from a blood product.

0065Of the Term "isolating" refers to separating a substance from a mixture containing more than one Component. For example, platelets are separated from whole blood. The product, which is isolated, does not necessarily refer to the complete separation of that product from the other components.

0066Of the Term "macropores" generally means, that the diameter of the pores is greater than about 500 Å. Of the Term micropores refers to pores with diameters of less than about 20 Å. The term mesopores refers to pores with diameters of greater than about 20 Å, and less than about 500 Å.

0067Of the Term "macroporous" is used to describe a porous Structure with a considerable Number of pores having diameters of greater than about 500 Å is used.

0068Of the Term "macroreticular" is a relative Term that means that the structure has a high physical porosity (ie a big Number of pores is present) a porous adsorbent structure comprising, both macropores and micropores has.

0069Of the Term "mesh enclosure," "mesh pouch" and the like refers to an envelope, bag, Bag or the like, which are prepared with multiple pores. For example, attracts present invention provides a bag in consideration of the immobilized containing adsorbent, with pores of a size that the Blood product as the Kon taktieren the immobilized adsorbent allow, but the immobilized adsorbent particles within the Bag restrain.

0070Of the Term "separating element" refers to any type of device or element that a whole into sections or parts separate or may split. For example, in the present invention provides the use of a separating element to Separating a blood bag adapted to receive a blood product is drawn in two parts into consideration. Busy The blood product a portion of the bag before and during the treatment, whereas the other part of the adsorption resin occupied. In one embodiment, the separating member after the treatment of the blood product removed (eg. B. integrity of the partition member changes), whereby the treated blood product that coming into contact with the Adsorption resin is made possible. The separating element can be positioned either inside the bag or on its exterior be. When using the term "separating element", the term "removed" means that the insulation the two parts of the blood bag no longer exists; he means not necessarily mean that the separating element is not more with the Bag is connected in any way.

0071Of the Term "photoproduct" refers to Products resulting from the photochemical reaction that a psoralen accomplished when exposed to ultraviolet light irradiation becomes.

0072Of the Term "polyaromatic Compounds "refers located on polymeric compounds that contain aromatic groups in the backbone such as. for example, polyethylene terephthalate, or as pendant groups, such. as polystyrene, or both.

0073Of the Term "polystyrene network" refers broadly to to polymers of styrene (C₆H₅CH = CH₂) Monomers include: The polymers can be linear, consisting of a single covalent alkane chain be cross-linked with phenyl substituents, or, in general, with m- or p-phenylene radicals, or another bifunctional or hyper crosslinked structure to a two-dimensional polymer backbone to form.

0074Of the Term "psoralen removal method" refers to a substance or element for the removal of more than about 70% of the psoralen from z. B. a blood product is capable of, preferably more than about 90%, most preferably more than about 99%. A psoralen removal method can also remove other components of the blood product, such as Psoralen photoproducts.

0075Of the Expression "Reduce the concentration "relates to the removal of a portion of the aromatic compounds from the aqueous Solution, wherein the reduction of the concentration is preferably in the order of more than about 70%, more preferably in the order of about 90%, and most preferably in the order is of about 99%.

0076Of the Set "Distance of substantially the total quantity or a part of a small organic compound (z. B. a psoralen, psoralen derivative, isopsoralen, Acridine, acridine derivative or dye), the freely soluble "preferably refers present to the removal of more than about 80% of free in solution Compound, preferably, the distance of more than about 85%, even by preferably greater than about 90%, and most preferably the removal of more than about 99%.

0077Of the Term "resin" refers to a solid support (Such as particles or beads etc.), to interact with and bind to different small organic compounds, including psoralens, in a solution or liquid (Z. B. a blood product) capable is, while reducing the concentration of those elements in solution. The removal process is not limited to a particular mechanism limited. To the Example, a psoralen by hydrophobic or ionic interaction (Ie, affinity interaction) be removed. The term "adsorption" refers broadly to both natural organic substances and synthetic substances, as well as Mixtures thereof.

0078Of the Term "shaker" refers to any type of device for the thorough mixing of a blood product, such as a platelet concentrate is capable. The device may have a timing mechanism to the restriction to allow the mixing process to a particular duration.

0079Of the Term "sintering medium" refers to a structure by applying heat and pressure to a porous plastic is formed, including For example, a powdery thermoplastic polymer. Porous plastics can by mixing powders of polymers having relatively low melting point and then heating to be prepared, so that the plastic particles partially melt, but still a passageway for fluids to penetrate into the porous Mass leave. Sintered Adsorbensmedien can similarly Manner by absorption of carbon or other highly or non-melting adsorbent in the powder to be heated be prepared with a low melting point. Methods for generating the porous Plastic materials are described in US Patent Nos. 3,975,481, 4,110,391, 4,460,530, 4,880,843 and 4,925,880. The process causes the melting of the powder particles, resulting in the formation of a porous solid Structure results. The sintering medium may be formed into a variety of forms, by dissolving the polymer powder during the sintering process is introduced into a mold. The adsorbent can in the sintering medium by mixing the adsorbent particles with the powdered thermoplastic introduced polymer and then the sintering process are subjected.

0080Of the Term "stabilizer" refers to a compound or composition of certain of optimizing the adsorptivity resins capable is. Generally speaking, should suitable stabilizers in water and ethanol (or other wetting agents) be soluble to water and ethanol nonvolatile and safe for Be the transfusion in small amounts. Examples of stabilizers include, without limited to be, glycerol and low molecular weight PEGs. A "wetting agent" is a "stabilizer" distinguishable, that is assumed from the former that it adsorbent pores of those resins reopens, which are not cross-linked hyper (ie non-macroreticular resins). Wetting agent are not generally prevent pores under dry conditions shrivel, whereas stabilitylisa factors will prevent this. A general discussion the wetting and the wetting agent is shown in U.S. Patent No. 5,501,795 to Pall et al.

0081Of the Set "a desired biological activity maintained the blood product substantially "refers essential to maintaining the properties of a blood product, the for the potential performance of the product in a therapeutic measure for indicative being held. For example, where red blood cells are affected, is the in vivo activity not destroyed or substantially reduced when the ATP-ups, the extracellular potassium exit, the% hemolysis in the red blood cells, Policies have been treated by the herein compared be maintained to untreated samples substantially. For example, the change the amount of ATP between the treated red blood cells and the untreated red blood cells less than about 10% be. The change hemolysis between the treated red blood cells and the untreated red blood corpuscle after the storage can be less than about 1%, preferably be less than about 0.8%. The change in extracellular potassium exit between the treated and the untreated red blood cells red blood cells may be less than about 15%. In the case of plasma is the in vivo activity not destroyed or substantially reduced, when the amount of coagulation factors, z. B. factors I, II, V, VII, X, XI, or the change PT and PTT time when methods described herein by means of treated compared to untreated samples substantially maintained in the plasma remains. For example, the change in the amount of coagulation factors, such as, for. example, factors I, II, V, VII, X, XI between the treated and the untreated plasma Plasma less than about 20%, respectively; preferably less than about 10%. The change PT and PTT time for the treated plasma compared to untreated plasma can z. B. less than about 3 seconds and larger than 1 second, preferably 1.5 seconds. Regarding the platelets, then the in vivo activity not destroyed or substantially reduced if, for. example, the platelet yield, the pH of the aggregation reaction, the change in shape, the GMP-140, the morphology or hyotonische shock reaction in treated by the methods described herein red blood corpuscle maintained in comparison to untreated samples substantially will. It is further contemplated that the phrase "substantially maintained" for each of the Properties described in connection with a blood product values ​​may also comprise, for the Professionals of the area acceptable and as described in the literature are, including z. B. in Klein HG, ed. Standards for Blood Banks and Transfusion Services, 17th Edition, Bethesda, MD. American Association of Blood Banks., 1996

0082Of the Term "thiazine dyes" includes dyes, the sulfur atom contained in one or more rings. Of the common Thiazine dye methylene blue [3,7-bis (dimethylamino) phenothiazine-5-ium chloride). Other thiazine dyes include, without limitation to Azure A, azure C and thionine as described for. Example, in US Pat. No. 5,571,666 to Schinazi.

0083Of the Term "xanthene dyes" refers to Dyes which are derivatives of the compound xanthene are. The xanthene dyes can be divided into one of three broad categories: i) fluorenes aminoxanthenes or, ii) the rhodols or Aminohydroxyxanthene, and iii) the fluorones or Hydroxyxanthene. Examples of xanthene dyes that for Use with the present invention are contemplated include Rose bengal and eosin Y; These dyes are commercially obtainable from a number of sources (eg., Sigma Chemical CO., St. Louis, MI), and such. As described in US Pat. No. 5,571,666 to Schinazi.

adsorbent

0084Provided be adsorbent that of a member to reduce Concentration of small organic compounds in a blood product while substantially maintaining a desired biological activity of the blood product useful are.

0085the adsorbent can from any regular or irregular shape be offered for inclusion in the inert matrix, but preferably approximately spherical. If the IAD used with a flow-through member, so, the Particles have a diameter of greater than about 10 microns, preferably the particles have a diameter of between about 10 microns and about 200 .mu.m; preferably the particles have a diameter of between about 10 microns and about 100 .mu.m; most preferably, the particles have a diameter of between 10 microns and about 50 microns on. If the IAD used with a charge element, so wei sen the particles have a diameter of greater than about 300 microns and less about 1200 microns on; preferably, the particles have a diameter of between about 300 microns and about 1200 microns on.

0086On high surface area is for the particle characteristic. Preferably, the particles a surface area of between about 750 m²/ G and about 3000 m²/ G. More preferably, the particles have a surface area of between about 1000 m²/ G and about 3000 m²/ G. Most preferably, the particles a surface area of between about 1750 m²/ G and about 3000 m²/ G.

0087For the use in the element of the present invention, suitable adsorbent can be made of any material on which the small organic molecules be adsorbed, with the restriction that the material of the biological activity a liquid do not substantially impair the contact back. The adsorbent can for example, materials such as activated carbon, hydrophobic resins or ion exchange resins exist.

0088at a preferred embodiment, pass the adsorbent particles of activated carbon, of either natural or synthetic sources derived. Preferably, the activated carbon comes from synthetic sources. Non-limiting examples of activated carbon include: Picatiff Medicinal, which is available from PICA USA Inc. (Columbus, OH), Norit ROX 0.8, which is available from Norit Americas, Inc. (Atlanta, GA), Ambersorb 572, which is obtainable from Rohm &amp; Haas (Philadelphia, PA) and G-277, which are available is of PICA (Columbus, OH).

0089In a preferred embodiment, , the particles in Norit A Supra, which is available from Norit Americas, Inc. (Atlanta, GA). Norit A Supra is an activated carbon in USP grade, which is formed by steam activation of lignite. The activated carbon has a very high total surface area (2000 m²/ G) and is very microporous nature.

0090In another preferred embodiment, can the particles may be hydrophobic resins. Non-limiting examples of the hydrophobic Resins include the following polyaromatic adsorbents: Amberlite®^®-Adsorbenzien (Z. B. Amberlite^® XAD-2, XAD-4 and XAD-16) available from Rohm &amp; Haas (Philadelphia, PA); Amber chrome^®-Adsorbenzien, Available from Toso Haas (TosoHaas, Montgomeryville, PA); Diaion^®/ Sepabeads^®-Adsorbenzien (Z. B. Diaion^® HP20) available from Mitsubishi Chemical America, Inc. (White Plains, NY); Hypersol-Macronet^®-Sorbentharze (Z. B. Hypersol-Macronet^®-Sorbentharze MN-200, MN-150 and MN-400), available from Purolite (Bala Cynwyd, PA); and Dowex^®-Adsorbenzien (Z. B. Dowex^® XUS-40323, XUS-43493 and XUS-40285), available from Dow Chemical Company (Midland, MI).

0091Preferred Particles are hydrophobic resins polyaromatic adsorbents are a hyper-linked polystyrene network, such as Dowex^® XUS-43493 (Known commercially as Optipore^® L493 or V493) and Purolite MN-200th

0092Hyper Networked Polystyrene networks, such. As Dowex^® XUS-43493 and Purolite MN-200, consisting of non-ionic macroporous and macroreticular resins. The non-ionic macroreticular and macroporous Dowex^® XUS-43493 has a high affinity for psoralens on, including for example, 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, and exhibits excellent wetting properties. The term "superior wetting properties" means that dry (That is, essentially anhydrous) adsorbent does not with a wetting agent wetted (eg., ethanol) prior to combining with the blood product must be in order for the adsorbent, the concentration of small organic Compounds in the blood product effectively reduced.

0093Hyper Networked Polystyrene networks such as Dowex^® XUS-43493 and Purolite MN-200 are in the form of spherical Particles having a diameter ranging from about 100 microns to about about 1200 microns to prefer. For Flow elements, the particles have a diameter in the range from about 10 to about 100 microns on. For Charge elements, the particles have a diameter in the range from about 100 to about 1200 microns on. Adsorbent particles, including, for example Dowex^® XUS-43493, preferably have extremely high internal surface areas and relatively small Pore ​​(z. B. 46 Å) on. The inner surface area the particles may be from about 300 to about 1100 m²/G Betra gen; preferably 1100 m²/G. The pore size of the particles can be more than 25 Å and be less than 800 Å; preferably about 25 Å to about 150 Å; most preferably from about 25 Å to about 500 Å. Although the present invention should not be limited to the mechanism by which effected the reduction of small organic compounds to, but is a hydrophobic interaction as the primary mechanism adsorption considered. Its porous nature confers the adsorption Selectivity, by small molecules access to a larger share the surface area relative to larger molecules (d. h. proteins) and cells allows becomes. Purolite^® has many similar Features such as Dowex^® XUS-43493 on such as a high affinity for psoralens and excellent wetting properties, and therefore is also preferred adsorbent is.

0094Poylstyrolpartikel can starting from its synthesis mechanism and the physical and functional Properties are classified as i) conventional networks and ii) hyper-linked networks. Preferred adsorbents have a high surface area on, have pores that do not shrink, do not require Wetting and have for Materials, the red blood cells or platelets include, in addition extremely small amounts of small particles and foreign particles (eg. As dust, fibers, non-adsorbing particles and unidentified Particles) on. For a batch element preferably have the small particles a Diameter of less than 30 microns on. For a flow element preferably have the small particles a Diameter of less than 5 microns on. also adsorbents, the preferred low levels of extractable remaining monomer, crosslinkers, and other organic extractable Substances.

0095the usual Networks are primarily Styrene-divinylbenzene copolymers in which divinylbenzene (DVB) as a Crosslinking agent is used (ie, the agent, the linear polystyrene chains linked together). These include polymer networks the "gel-like" polymers. The gel-like Polymers are homogeneous, non-porous styrene-DVB copolymers, as by obtained copolymerization of monomers. The macroporous adsorbents represent a second class of conventional networks. They are obtained by copolymerization of monomers in the presence of diluents obtain that precipitate the growing polystyrene chain. The formed by this method Polystyrene network has a relatively large inner surface area (Up to hundreds of square meters per gram of polymer); Amberlite^® XAD-4 is produced by such a method.

0096in the provide contrast to the conventional networks described above The preferred adsorbents of the present invention (eg. B. Dowex^® XUS-43493) hyper-linked networks. These networks are through networking linear polystyrene chains either in solution or in a swollen State generated with bifunctional agents; the preferred bifunctional Agents produce conformational restricted crosslinking bridges, which is assumed that on the pore shrinkage prevent, when the adsorbent in a substantially anhydrous (Ie, "dry") state.

0097The hyper-connected networks are attributed to three primary characteristics, they of the conventional Networks differ. First, the polymer chains are in low Density before due to the bridges, that hold the polystyrene chains apart. As a result, the Adsorbents generally a relatively large porous surface area and pore diameter on. Second, the networks are swellable; ie, the volume of the polymer phase increases upon contact with organic molecules. After all The hyper-linked polymers "stretched" in the dry state, that is, the rigidity of the network in the dry state prevents the chain-to-chain attractions. relaxes the strain However, when the adsorbent is wetted, which swellability the network in liquid Media increases. Davankov and Tsyurupa, Reactive Polymers 13: 27-42 (1990); Tsyurupa et al., Reactive Polymers 25: 69-78 (1995).

0098Several Crosslinking agent were to produce bridges between polystyrene chains successfully applied, including p-Xylendichlorid (XDC) Monochlorodimethyl (MCDE), 1,4-bis-Chlormethyldiphenyl (CMPD) 4,4'-bis (chloromethyl) biphenyl (CMB), dimethylformal (DMF), p, p'-bis-chloromethyl-1,4-diphenylbutane (DPB) and tris- (chloromethyl) -mesitylene (CMM). The bridges between polystyrene chains by reacting one of these cross-linking agent with the Styrolphenylringen formed by means of a Friedel-Crafts reaction. To link the resulting bridges Styrene phenol rings present on two different polystyrene chains are. See, eg., US Pat. No. 3,729,457.

0099the bridges are especially important, they generally turn off the requirement of a "wetting agent". The called, that the bridges a shrinkage of the pores prevent, when the adsorbent in a substantially anhydrous (ie, "dry") state, which is why they do not with a wetting agent prior to contacting the adsorbent with a blood product "again must be opened ". To to prevent shrinkage of pores, conformational restricted bridges should formed will. Some bifunctional agents like DPB found no general limited Conformation: for example, contains DPB four successive methylene units for conformational rearrangements prone are. Therefore DPB is not a preferred bifunctional agent for Use is in the present invention.

0100Some the structurally-related characteristics of the above-described Adsorbent are summarized in Table A.

TABLE A <img img-content="tb" img-format="tif" he="144" wi="167" file="00250001.tif" />

Processing the adsorbent

0101the adsorbent can for the removal of fine particles, salts, potential extractables and endotoxins are further processed. The removal of these extractable components is typically through treatment with either organic solvents, Steam or supercritical fluids made. Preferably, the particles are sterilized.

0102Several Companies currently distribute "purified" (ie processed) Versions of commercial Adsorbent. about test the processing of the adsorbent particles (eg. as resins) addition these companies the adsorbents, wherein the finished adsorbent be sterile certified (USP XXI), pyrogen-free (LAL) and free of detectable extractables (DVB and Gesamtorganik) are.

0103the heat processing (Z. B. steam) is an effective method of treatment of adsorbent represent. F. Rodriguez, Principles of Polymer Systems, (Hemisphere Publishing Corp.), p 449-53 (3rd ed., 1989). Supelco, Inc. (Bellefonte, PA) uses a proprietary thermal process without solvent, the Dowex^® XUS-43493- to clean and Amberlite adsorbents. The main advantage of Use of steam is that this the adsorbent no potentially extractable adds. However, a large disadvantage is that this process can remove the pores of the resin beads of water; effective performance of some adsorbents requires that the beads rehydrated prior to contacting with the irradiated blood product will.

0104On Advantage of the cleaned / processed adsorbent is its extremely small amount of particles with diameters of less than than 30 microns. A preliminary test of adsorbents (Dowex^® XUS-43493 and onberlite^® XAD-16) which were processed by Supelco was used to determine the particle counts made. The results of these tests indicated that foreign particles (Eg. As dust, fibers, non-adsorbent and unidentified Particles) were not available and that fine particles (less than 30 microns) substantially were absent.

Use of the wetting agent and stabilizers with adsorption resins

0105It can Methods to prevent desiccation and loss of adsorption capacity of Particles are applied, such as with Amberlite^®, the a portion of its adsorption capacity under certain conditions loses (z. B. drying).

0106at a method can the particles, materials or elements are produced in a wet state, the sealed and unable is to dry out. This method is more important disadvantages connected. The shelf life of products could be reduced because the amounts of extractables from the materials with the time could increase. Sterilization could limited to a process Dampfpro be because a gamma irradiation of moist polymers usually is not made. The production of an element which requires moist attitude of a component, is generally more difficult as the preparation of a dry element; For example, biological Stress and endotoxins a cause for concern, when a long time delay between the composition of the element and the terminal sterilization lies.

0107A second method of contraception a loss of adsorption capacity refers to the use an adsorbent one, which does not adversely by drying is affected. macroreticular As previously indicated, require Adsorbents highly crosslinked porous structures have (eg. B. Dowex^® XUS-43493 and Purolite^® MN-200), generally no wetting agent because the cross-links a Shrinkage of pores prevent. Unlike Amberlite^® XAD-16, keep these macroreticular adsorbents a very high proportion their original activity in the dried state.

0108at A third method, a loss of adsorption capacity on drying out by hydrating Amberlite®^® XAD-16 and related adsorbents (z. B. Amberlite^® XAD-4) in the presence of a non-volatile Wetting agent can be prevented. For example, in Using Amberlite^® XAD-16 as the adsorbent the Adsorbenskügelchen prior to use during handling, sterilization and storage partially dry. If the Water content of these adsorbents below a critical level, so enters rapid loss of adsorptive a (possibly due to the "Together shrinkage" of the pores); therefore have to for a optimum effectiveness, the pores are "opened" prior to use with a wetting agent.

0109stabilizers are to maintain the adsorption capacity near its maximum effect when specific adsorption resins drying conditions are subjected. It is believed that the use of stabilizers of preventing an Together shrinkage of the adsorbent pores serves.

0110On acceptable stabilizer should be soluble in water and ethanol, opposite to Ethanol and water non-volatile and represent be sure transfusion in small amounts. Glycerol and low molecular weight polyethylene glycol (Z. B. PEG-200 and PEG-400) are examples of stabilizers represents that possess these properties. Glycerol has a positive Hämokompatibilitäts history on. It is often the blood as a Cryokonservierungsmittel in frozen storage red blood cell preparations added. See, e.g., Chaplin et al, Transfusion 26:.. 341-45 (1986); Valeri et al., Am. J. Vet. Res. 42 (9) 1590-94 (1981). Solutions, the contain up to 1% glycerol, are transfused routinely wherein glycerol solutions commercially available are (z. B. Glycerolite 57 solution Fenwal Laboratories, Deerfield, IL). Adsorbenskügelchen as Amberlite^® XAD-16 can be stabilized in ethanol and glycerol.

0111low molecular weight Polyethylene glycols, such as frequently used as pharmaceutical raw materials, can also be used as stabilizers be used. PEGs provide liquid and solid polymers of general chemical formula H (OCH₂CH₂)_nOH represents wherein n greater than or equal to 4. The PEG formulations is usually followed by a number that its average molecular weight equivalent; has, for example PEG-200 has a molecular weight of 200 and a molecular weight range of 190-210 on. PEGs are available in a number of formulations available commercially (eg. B. Carbowax, Poly-G and Solbase).

Inert matrices for Partikelimmobilisation

0112the Adsorbent particles are immobilized by an inert matrix. The inert matrix can be made of a synthetic or natural be prepared polymer. For example, the inert matrix, a synthetic or natural be polymer fibers, such as a fiber network. The inert matrix may consist of sintered polymers. The inert matrix is ​​also as the other components of the element, preferably biocompatible and affects the biological activity of a material on contact essentially not.

0113At the preferred are the synthetic fibers are polyethylene fibers (Air Quality Filtration (AQF), a division of Hoechst Celanese (Charlotte, NC)). Further preferred examples of the synthetic fibers are polyolefin, Polyester or polyamide fibers. To know more advanced examples of synthetic fibers include Polypropylene, polyvinyl alcohol and polysulfone.

0114at a preferred embodiment, comprising the synthetic polymer fiber comprises a first polymer core with a high melting point surrounded by a sheath with a lower melting point. The polymer core may be a Polyethylenpolyesterterephthalat core. The sleeve , a nylon sleeve or may be a modified polyester sheath. The fibers are commercially available from Unitika (Osaka, Japan) and Hoechst Trevira GmbH &amp; Co. (Augsburg, Germany).

0115To examples of natural Polymer fibers include Cellulose fibers derived from a variety of sources, such as about jute, Kozu-paper, kraft pulp and Manila hemp. networks synthetic or nat ürlichen Polymer fibers have been used for the production of filters, such as described in US Patent Nos. 4,599,145 and 5,639,376.

0116For the construction of sintered particles of suitable synthetic polymers are polyethylene a high density, ultra high molecular weight polyethylene, polypropylene, Polyvinyl fluoride, polytetrafluoroethylene, nylon 6. More preferred are the sintered particles polyolefins such as polyethylene.

0117fibers without adsorbent are for The present invention contemplated, such as fibers, preferably a large, porous Adsorptionsoberflächenbereich include or other adsorptive circumstances, the reduction in a the concentration of small organic compounds easier.

Immobilization of the particles

0118In one embodiment the adsorbent particles are immobilized by an inert matrix to an adsorption medium for reducing the concentration of small to produce organic compounds in a material. The inert Matrix may be a three-dimensional network, including a synthetic or natural Polymer fiber network with adsorbent particles immobilized therein.

0119Preferably the adsorbent comprises small porous adsorbent with highly porous Structures and very large Inner surface regions, as described above, the immobilized by an inert matrix are. When a biological material in contact with the adsorption medium brought so impaired the adsorption medium, the biological activity or other properties the material preferably only insignificantly.

0120the Technology for the immobilization of Adsorbenskügelchen on a fiber network for the construction of air filters has been described in U.S. Patent No. 5,486,410 and US. Pat. No. 5,605,746. As in<figref idrefs="S117">1</figref> shown, pass the polymer fibers <figref>600</figref> of the fiber network from a polymer core <figref>602</figref> (Eg., Polyethylene terephthalate (PET) with a high melting point surrounded by a polymer sleeve <figref>604</figref> (Z. For example, nylon) having a relatively low melting point. See US Patent No. 5,190,657 to Heagle et al. The fibrous resin is prepared, by first the Adsorbenskügelchen uniformly in the fiber network be distributed. Next The network is rapidly heated (eg., 180 ° C × 1 min), which melts the the polymer sleeve fibers <figref>600</figref> and the binding of the Adsorbenskügelchen <figref>606</figref> and other fibers which form a cross-linked fiber network, causes as shown in <figref idrefs="S118">2</figref>, As in<figref idrefs="S119">3</figref> and <figref idrefs="S120">4</figref> illustrated (Not to scale) included, the fiber networks generally said three layers; two outer layers<figref>607</figref>. with fibers <figref>600</figref> are tightly packed, and a less tight inner layer <figref>609</figref>That the Adsorbenskügelchen <figref>606</figref> and fewer fibers <figref>600</figref> contains. In a preferred embodiment can the edges of the fibrous Resin to be sealed with polyurethane or other polymers. alternative can, as in <figref idrefs="S119">3</figref> and <figref idrefs="S120">4</figref> shown, heat seals 608 in the resulting fibrous made resin at predetermined intervals will; the heat seals can For example, in the fibrous Resin are produced in the pattern shape of squares. Thereupon the fibrous Resin by the heat seals to obtain resin samples are cut, the desired mass (Z. B. preferably less than 5.0 g, and more preferably less than 3.0 g) at Adsorbenskügelchen contained and have a size, which for placement within a blood product container suitable is. The heat seals serve fraying of the cut fibrous resin to prevent and in immobilizing the Adsorbenskügelchen helpful. However, the use of such heat seals is at Execution of the present invention is not required. In an alternative embodiment , as in <figref idrefs="S120">4</figref> illustrated, the Adsorbenskügelchen <figref>606</figref> not attached to the fibers themselves, but rather between the denser outer layers <figref>607</figref> of the Fibers and with the heat seals <figref>608</figref> immobilized; this embodiment may also samples of fibrous Media arise that a defined amount of adsorbent according to the Cutting through the heat seals contain.

0121For the present Invention is also the use of a bonding agent (eg. as an adhesive) for fixing the adsorbent resin drawn to the fibers into consideration. Although a chemical bond to the Adsorbenskügelchen the fiber network preferred, but the beads about that are also physically trapped within the fiber network; This can, for example, by surrounding the beads with sufficient fibers be achieved so that the beads are held in position.

0122Other Ways of making the adsorbent particles immobilized in a fiber network can be, are also contemplated. The particles may under Application of a dry process are immobilized as described in US Patent Nos. 5,605,746 and 5,486,410 (AQF-patents). The particles can using a Naßlegeprozesses be immobilized, as described in US Patent Nos. 4,559,145 and 4,309,247. The particles may be immobilized using a wet method, as described in US Pat. No. 5,616,254, which is incorporated herein by reference is. Where a wet method for constructing a matrix of natural Polymer fibers is applied, the inert matrix contains preferably a binder for adhering the adsorbent particles to the fibers. nonlimiting Examples of binders include melamine, polyamines and polyamides. The matrix contains typically 1% or less of such binders.

0123There, where the inert matrix of particles of synthetic polymers designed is, which are sintered with particles of adsorbent, it is important that the adsorbent particles have a higher melting temperature than the matrix.

0124Preferably includes the resultant adsorption medium known amounts of adsorbent per surface. For batch items makes the adsorbent per area about 100 g / m² to about 500 g / m² out, preferably about 250 g / m² to about 350 g / m², For Flow elements makes the adsorbent per area is about 300 g / m² to about 1100 g / m² from, preferably about 500 g / m² to about 700 g / m², Therefore leaves the appropriate amount of adsorbent for a specific purpose it is considered simply by cutting a predetermined area of the fibrous Resin measure (ie there is no weighing of the fibrous resin).

0125The Adsorption is preferably biocompatible (ie it generates no toxic, harmful or immunological reaction); has a minimal effect on the Properties of the material, such as the blood product (eg. As platelets and Clotting factors) and is non-extractable with toxic materials connected. The immobilized adsorbent particles of the adsorption preferably have a high mechanical stability (d. h. No fine particle generation). The adsorption of a batch element contains about 25-85% Adsorbent by weight, preferably about 50-80% adsorbent at a loading of about 100-500 g / m², More preferably about 50-80% adsorbent at a loading of about 250-350 g / m²,

0126The adsorption for includes a flow element about 20-70 Wt .-% adsorbent, preferably 30-50 wt .-%. Preferably, the adsorbent contains about 30 wt .-% of the adsorbent when a fibrous matrix is used. Where the particulate Sintered matrix used with ground polymeric adsorbent is, contains the adsorption medium is preferably about 50 wt .-% of the adsorbent particles.

Coating the adsorbent

0127the Surface hemocompatibility of the particles, Matrices or adsorption, by coating their surfaces with be improved a hydrophilic polymer. Examples of the hydrophilicpolymers count Poly (2-hydroxyethylmethacrylate) (pHEMA), which can be obtained for example, from Scientific Polymer Products, Inc. (Ontario, NY), and cellulose-based polymers, for. example, ethyl cellulose, which can be obtained from Dow Chemical (Midland, MI). See z. B. Andrade et al., Trans Amer. Soc. Artif. Int. Organ XVII: 222-28 (1971). Other examples of coatings include polyethylene glycol and Polyethylene oxide, as also available from Scientific Polymer Products, Inc. The polymeric coating can increase hemocompatibility and the risk of the generation of small particles due to mechanical Elimination diminish.

0128the adsorbent surface can also be modified with immobilized heparin. Furthermore can strong anion-exchanging-Polystyrenedivinylbenzene adsorbents via a Heparin adsorption be modified. Heparin, a polyanion adsorbed very strongly to the surfaces of adsorbents which have strong anion exchanging properties. A variety of quaternary Amine-modified polystyrene divinylbenzene adsorbents is commercially available.

0129the Coating can using a variety of methods be applied, including radio frequency glow discharge, as described in US Pat. No. 5,455,040, and the Wurster coating process (Carried out by International Processing Corp. (Winchester, KY)).

0130In one embodiment , the Wurster coating process by suspending the adsorbent (On generally Air pressure) are carried out in a chamber, so that the hydrophilic Polymer z. B. pHEMA, evenly on all surfaces sprayed the adsorbent can be. As illustrated in Example 3, showed Dowex^® XUS-43493 with uniform spraying with pHEMA increased platelet yield as well as a tremendous effect on the change in shape of the platelets with increasing amounts of coating. It has been found that the Wurster coating process, the outer surface of the selectively Adsorbensfläche coated, whereas the inner porous surface RECOURSE virtually unaffected.

0131In a preferred embodiment, the coating by immersing the immobilized adsorption medium be applied in the hydrophilic polymer (see Example 3). This process is simpler and less expensive than spraying the Adsorbent particles with the hydrophilic polymer.

0132Of the Process is not limited to a process in which the coating the adsorption medium is applied at a certain time. For example, in one embodiment the pHEMA coating after the production of the adsorption medium, but before the heat sealing the adsorption medium applied. In another embodiment, is the adsorption initially heat sealed and then applied the pHEMA coating. In addition to the coating of the Adsorption medium is used connected to the pHEMA application rinsing to remove loose particles and fibers.

0133With increasing amount of coating it is for some small organic Compounds more difficult to pass through the coating in order to the particle surface to arrive, leading to a decrease in the adsorption kinetics. So must generally with increasing amount of coating increased mass be used on the adsorbent to achieve the same removal kinetics to achieve as coated adsorbent. In one embodiment, is the optimum amount of pHEMA coating the minimum amount of coating, wherein a protective effect on platelet yield and in vitro platelet function is observed (0.1-0.5%).

0134the coatings can sensitive to its sterilization. For example, a gamma sterilization lead to cross-linking and / or columns of the coating. Therefore the type (E-beam gg. gamma irradiation) and dose of sterilization affect the properties of the coated adsorbent. As a general rule is preferably an E-beam sterilization.

elements

0135It are elements for reducing the concentration of small organic Compounds in materials such as blood products provided. The Element can, for example, a flow element or a batch element be. Examples of flow and Charge elements are in <figref idrefs="S133">16</figref> shown. Flow and batch elements are known in the literature and for example, in PCT Publication WO 96/40857 described.

0136the Charge elements of the invention comprise a container, such as a blood bag, of the containing adsorption media with the immobilized particles. In a embodiment is a blood product placed in a blood bag, the adsorption medium, the contains, and the bag for shaken a specified period.

0137To the For example, in one embodiment an adsorption z. B. immobilized Dowex^® XUS-43493, in a blood product container placed (z. B. a PL 146 plastic containers (BaxterHealthcare Corp. (Deerfield, IL)), either on a platelet shaker (Helmer Laboratories (Novesvill, IN)) or a rotary device (Helmer Laboratories (Novesvill, IN)) for about 24 hours, is held at room temperature and under different Temperature conditions is stored. The size of the blood product container can vary from about 600 to about 1000 ml. The storage temperature may about 4 ° C to about 22 ° C be.

0138Methods to reduce the presence of particles of adsorbent, which is possibly from Adsorption replace, can be applied.

0139For the present Invention, a batch element contemplated that the immobilized adsorption medium, retained in a container such as a mesh bag / Bag contains. The mesh bag / Bag may consist of a woven, non-woven or membrane-like container material be constructed. In one embodiment, , the woven mesh bag made of a polyester or nylon in medical quality be constructed. The preferred embodiment is polyester. To commercial membranes count limited without it to be, Supor^® 200, 800, 1200 - hydrophilic Polyethylensulfonat- (PES) membranes (Gelman Sciences (Ann Arbor, MI)); Durapore^® - Hydrophilic modified polyvinylidene fluoride (PVDF) (Mantee America Corp. (San Diego, CA)) and hydrophilic modified polysulfone membranes with integrated hydrophobic vents for z. B. Gemini membranes (Millipore (Marlborough, MA)); and membranes that polycarbonate with a polyvinylidene coating include (Poretics (Livermore, CA)). The containers may, after the addition of the adsorption medium be sterilized.

0140Flow elements allow the reduction of the concentration of small organic Compounds from materials such as blood products by perfusing the blood product through the flow element.

0141Where the element is a flow element, is the adsorption preferably about 3 to 30 mm thick to an even flow the biological fluid without promoting significant pressure drop. Preferably, the Medium about 3 to 15 mm thick. More preferably, the medium comprises about 5 to 8 mm thick. Where the element is a charge element, the adsorbent medium may be about 2 to 30 mm thick. Preferably the medium is about 2 to 10 mm thick.

Preferred embodiment for platelets

0142Preferably is a batch or flow-through element for reducing the concentration of small organic compounds in a material comprising platelets, used at substantially maintaining the biological activity of the platelets. A batch element is preferred. includes the adsorption to an inert matrix immobilized adsorbent. Preferred Particles are highly porous and have a surface area of greater than about 750 m²/ G.

0143Especially preferred particles are polyaromatic adsorbents hyper linked a include polystyrene network, such as Dowex^® XUS-43493 or Purolite MN-200th The preferred inert matrix includes a synthetic or natural polymer fiber. In a preferred embodiment contains the inert matrix is ​​a synthetic polymer fiber which first a Polymer core with a high melting point includes, surrounded by a sleeve a lower melting temperature. The polymer core, a polyethylene terephthalate core be. The sleeve , a nylon sleeve or may be a modified polyester sheath. are staple fibers commercially available from Unitika (Osaka, Japan) and Hoechst Trevira.

0144In one embodiment the batch element comprises an adsorption media, a particle retention element (Z. B. mesh, non-woven material, or membrane) and a housing (Z. B. blood storage container).

0145Examples of small organic compounds that, by means of the elements Materials and methods of the present invention are reduced, are psoralen, psoralen derivatives, Isopsoralene, psoralen photoactivation products, acridines and acridine derivatives.

0146the platelets are typically used within 3 days after delivery, can but be stored for up to 5 days at room temperature, so it would be advantageous platelets in contact with the adsorption medium for the entire storage period to leave. would preferably the procedure an acceptable platelet yield result (eg. B. less than 10% loss). One by the present invention contemplated method allows extended storage by improvement Hemocompatibility the adsorbent surface.

0147the Using an adsorption medium, which on an inert matrix comprising immobilized adsorbent particles, allows a reduction the concentration of small organic compounds without substantial Loss in platelet count. Of the Expression "without any substantial Loss " to a loss in platelet count of less than about 10%, preferably less than about 5%, over a period of 1 day preferably 5 days. Furthermore, the period over which the blood platelets with brought together the adsorption medium without substantial loss in platelet count can be, greater than the period of time over the platelet with only the adsorbent can be contacted. the Immobilization of the particles allows unexpectedly both extended Contact time and a reduced loss in platelet count. the platelets can not typically with non-immobilized adsorbent longer than about 20 hours without a substantial loss in platelet count, z. For example, about 80% yield, can be contacted. In contrast, the platelets can the adsorption, the immobilized on an inert matrix includes adsorbent, for more than 20 hours, eg., about 1 to 5 days, without a substantial Loss contacted in platelet count will.

0148It should also the in vitro platelet function (Z. B. shape change, GNP-140, pH) in platelets, which are stored in the presence of the adsorption medium, in Ver equal to platelets, to be stored without adsorption, seen over time better. Platelets stored in the presence of the adsorption medium can have a pH of greater than about 6 or smaller than 7.5.

Preferred embodiment for plasma

0149Preferably is a batch or flow-through element for reducing the concentration of small organic compounds in a material, the plasma comprises, in substantially maintaining the biological activity of the plasma used. A flow element is preferred. includes the adsorption to an inert matrix immobilized adsorbent. Preferred Particles are highly porous and have a surface area of greater than about 750 m²/ G.

0150Especially preferred particles are Norit A Supra, which are available from Norit Americas, Inc. (Atlanta, GA). Norit A Supra is an activated carbon in USP grade, which is formed by steam activation of lignite. This activated carbon has a very high total surface area (2000 m²/ G) and is very microporous nature.

0151Furthermore can selected the particles be of any of the following particles, wherein the particles preferably a diameter ranging from 10 microns to 100 microns, either by grinding, direct synthesis comprise or other method, and are activated carbons such. B. Picactif Medicinal (PICA USA, Columbus, OH), synthetic carbonaceous Adsorbents such as Ambersorb 572 (Rohm &amp; Haas, Philadelphia, PA), hydrophobic Resins such as Amberlite^®-Adsorbenzien (Z. B. Amberlite^® XAD-2, XAD-4 and XAD-16), available from Rohm &amp; Haas (Philadelphia, PA); Amber chrome^®-Adsorbenzien, Available from Toso Haas (TosoHaas, Montgomeryville, PA); Diaion^®/ Sepabeads^®-Adsorbenzien (Z. B. Diaion^® HP20) available from Mitsubishi Chemical America, Inc. (White Plains, NY); Hypersol-Macronet^®-Sorbentharze (Z. B. Hypersol-Macronet^®-Sorbentharze MN-150 and MN-400), available from Purollite (Bala Cynwyd, PA); and Dowex^®-Adsorbenzien (Z. B. Dowex^® XUS-40323, XUS-43493 and XUS-40285) available from Dow Chemical Company (Midland, MI).

0152the inert matrix can be a synthetic or natural polymer fiber. In a preferred embodiment, is the inert matrix of sintered particles or cellulose.

0153Examples the small organic compounds represented by the elements, materials and methods of the present invention to be reduced, are psoralens, Psoralen derivatives, Isopsoralene, psoralen photoactivation products, methylene blue, Phenothiazine, acridine.

0154There, where the biological fluid, the small molecules contains, the concentration of which is to reduce, plasma is the element preferably a flow-through element that maintains adequate levels of the clotting activity. the Measures of clotting activity include the prothrombin time, activated partial thromboplastin time and functional measurements of the coagulation factors I, II, V, VII, VIII, IX, X, XI and XII. adequate functional measure of clotting factor activity is greater than 80 the amount of wandering through the element, or in the case of Clotting times, one which remains in the normal range, as it detects each laboratory that performs this type of test. To preferred measures of clotting activity include PT and PTT, as these readings the insgesamten coagulability the plasma provides, and factors I, II, V, VII, X, XI and XII, since these factors usually not be replaced by recombinant proteins. It is preferred that more than 90% of the clotting activity of these factors relative to the amount is maintained before wandering through the element and changes are complied with PT and PTT of less than 1.5 seconds.

0155at one embodiment , the flow element and an adsorption include a housing. The housing should an even flow promote the plasma, in order to ensure a good utilization of the medium, and should allow the Plasma when filling enable, heart uschi just air in front of him, thereby turning a blistering, the contact area, the decrease between the plasma and the adsorption and thus would reduce the use of the medium. The housing may may be flat or a substantial have depth to an adsorption or particle retention medium is not flat, but for example, a cylindrically shaped Adsorption is to be incorporated therein. In a preferred embodiment , the housing flat. The housing can feeds and processes have in different orientations, for example feed above / drain top or inlet bottom / drain below. In a preferred embodiment is the flow at the bottom to a good drainage to promote, and the feed above to promote the utilization of the medium.

0156In Alternatively, may be a flow-through element, which comprises an adsorption medium and a housing, also a particle retention medium contain. In a preferred embodiment, the element contains a particle retention medium downstream the adsorption medium for retention of of Particles that have fallen from the adsorption, while Maintaining a high liquid flow rate and high recovery of proteins. The particle retention medium can be a membrane, one produced by dry or wet methods of matrix Fibers, a sintered polymer matrix, a fabric material, a non-woven Material (non-woven polyester) or a combination thereof be.

0157The housing holds the element the particle retention medium in an approximately parallel orientation downstream the adsorption medium. (US Pat. No. 5,660,731 describes examples of filter housings.) The housing may consist of a suitably rigid, impervious material designed be, that does not significantly impair the biological activity of a liquid. Preferably, the housing constructed from a synthetic polymer. Non-limiting examples such polymers include Poylacryl-, polyethylene, polypropylene, Polystyrene and polycarbonate plastics.

0158Where the element is a flow element, the adsorbent medium should containing the immobilized on an inert matrix particles, 3 to be 30 mm thick to a steady stream of biological fluid without promoting significant pressure loss. Preferably should the medium be 3 to 15 mm thick. Preferred should the media be 5 to 8 mm thick.

0159Where the element is a flow element is a gravity flow prefers. More preferably, the member is a gravity flow element, to enable the of flow rates from 10 to 80 ml / min with differential pressures of 12-72 Inches of water is constructed. Preferred allowed the element flow rates from 30 to 60 ml / min at a differential pressure of 24 to 48 inches of water.

Preferred embodiment for red blood corpuscle

0160Preferably is a batch or flow-through element for reducing the concentration of small organic compounds in a material which red blood corpuscle comprises, in the substantial retention of biological activity of the red blood corpuscle used. The adsorption medium comprises at an inert matrix immobilized adsorbent. Preferred particles are highly porous and have a surface area of greater than about 750 m²/ G.

0161Preferably is the element a flow or batch item, the biological, the activity of red blood cells by reducing the concentration of small organic compounds maintains substantially. The embodiment the flow element comprises two components: an adsorption and a housing, and optionally a particle retention medium. The embodiments the particle retention medium and housing are as described above for the embodiments the plasma element bewrote. In another embodiment, is the element of the red blood cells a batch item that is not the biological activity of a liquid on contact significantly impaired. The embodiment the batch element comprises an adsorption medium, which at a inert matrix immobilized particles and optionally a particle retainer contains.

0162In one embodiment the particles of activated carbon. Preferably, the activated carbon comes from synthetic sources. Non-limiting examples of activated carbon include Picactif Medicinal, which is available from PICA USA (Columbus, OH); Norit ROX 0.8, which is available from Norit Americas Inc. (Atlanta, GA); and G-277, which is available from Pica USA (Columbus, OH).

0163Where the element for the red blood cells a charge element is, the adsorbent preferably is of a a synthetic source derived activated carbon, such as Ambersorb 572. In the Ambersorbs is synthetic activated carbonaceous (z. B. carbon-rich) adsorbents from Rohm &amp; Haas (Philadel phia, PA) are produced. Ambersorbs are generally large spherical (300-900 microns) particles, which are more durable than typical activated carbons. The Ambersorbs by treating highly sulfonated porous polystyrene beads in a protected High-temperature activation process synthetically. These Adsorbents do not require pre-swelling, in order to achieve optimal adsorption.

0164In another preferred embodiment, can the particles may be hydrophobic resins. Non-limiting examples of the hydrophobic Resins include the following polyaromatic adsorbents: Amberlite®^®-Adsorbenzien (Z. B. Amberlite^® XAD-2, XAD-4 and XAD-16) available from Rohm &amp; Haas (Philadelphia, PA); Amber chrome^®-Adsorbenzien, Available from Toso Haas (TosoHaas, Montgomeryville, PA); and Diaion^®/ Sepabeads^®-Adsorbenzien (Z. B. Diaion^® HP20) available from Mitsubishi Chemical America, Inc. (White Plains, NY). In a particularly preferred embodiment, the particles Hypersol-Macronet^®-Sorbentharze (Z. B. Hypersol-Macronet^®-Sorbentharze MN-200, MN-150 and MN-400), available from Purolite (Bala Cynwyd, PA) or Dowex^®-Adsorbenzien (Z. B. Dowex^® XUS-43493 and XUS-40285), available from Dow Chemical Company (Midland, MI).

0165the includes preferred inert matrix a synthetic or natural Polymer fiber. In a preferred embodiment, the inert Matrix, a synthetic polymer fiber which includes a first polymer core comprises a high melting point surrounded by a sleeve a lower melting temperature. The polymer core, a polyethylene terephthalate his or polyester core. The sleeve , a nylon sleeve or may be a modified polyester sheath. The fibers are commercially available by Unitika (Osaka, Japan) and Hoechst Trevira (Augsburg, Germany).

0166In some embodiments is the adsorption in an envelope. In one embodiment, the charge element comprises an adsorption medium and a housing. In Alternatively, the charge element comprises an adsorption medium and a housing and can also a particle retention medium contain. In a preferred embodiment, the housing has a Blood bag with a volume of 600 ml to 1 liter. The particle retention medium a polyester woven fabric, a non-woven polyester or a membranous sheath include. Preferably, the batch element contacted by the red blood cells at 4 ° C or 22 ° C with shaking on a Period of 1 to 35 days.

0167the Using an adsorption medium, which on an inert matrix comprising immobilized adsorbent particles, allows a reduction the concentration of small organic compounds without substantial Loss of function of red blood cells. The term "without substantially Loss "refers to less than about 1% Change in hemolysis; preferably less than 0.8% change in hemolysis, greater than about 90% recovery red blood cells; preferably greater than 80% recovery of red blood cells, less than about 10% difference from the element-free control of the red blood corpuscle in terms of of change ATP concentration, and less than about 15% difference from the element-free Control of red blood cells in terms of of change extracellular Potassium concentration. On day 35, the change in the hemolysis is at least 10% higher for the IAD compared to the non-immobilized particles, preferably 20% higher and preferably 50% higher.

0168the Red blood cell function can using standard kits to be examined. In particular, the hemolysis can be obtained by measuring the absorbance at 540 nm of a supernatant sample of red blood cells in Drabkin's reagent (Sigma Chemical Company, St. Louis, MO) are determined. Potassium deficiency , by using a Na + / K + -Analysegeräts (Ciba-Corning Diagnostics, Medfield, MA) are examined. A quantitative enzymatic Determination of ATP in the whole red blood cellschen-samples using a standard kit (Sigma Diagnostics, St. Louis, MO) and measuring the absorbance at 340 nm in comparison with the water background possible.

0169Where to be cleaned biological fluid in red blood cells there, the element, the concentration of both the cyclic as well as acyclic small organic molecules in a sample of red blood corpuscle to reduce. Preferably, the element, the concentration of both acridine derivatives and thiols in a red blood sample to reduce. More preferably, the element concentration both of 5 - [(β-carboxyethyl) amino] acridine reduce and glutathione in a ro th blood sample. Standard HPLC assays can for determining the concentrations of 5 - [(β-carboxyethyl) amino] acridine and glutathione in the contacted with the element red blood cells be used. The mobile phase assay are 10 mM H₃PO₄ in HPLC grade water and 10 mM H₃PO₄ in acetonitrile. Zorbax SB-CN and YMC ODSAM-303 columns available from MacMod Analytical, Inc. (Chadds Ford, PA) and YMC, Inc. (Wilmington, NC).

applications

0170the present invention contemplates reducing the concentration of small organic compounds. For small organic compounds include Pathogen-inactivating agents such. As photoactivation products, Aminoacridines, organic dyes and phenothiazines. examples the pathogen inactivating agents include Furocumarine, z. B. psoralens and acridines. Following the treatment of a blood product with a pathogen-inactivating compound, such as described in US Patent numbers 5,459,030 and 5,559,250, the Concentration of the pathogen inactivating compounds in the blood product by contacting the treated blood product with an element the invention can be reduced.

0171In one embodiment the present invention contemplates a method for the inactivation of Pathogens in solution into consideration, the method comprising: a) providing, in any Direction: i) a cyclic compound, ii) a solution in the contamination with the pathogen is suspected, and iii) a fibrous resin; b) treating the solution with the cyclic compound to a treated product solution to obtain, wherein the pathogens are inactivated; and c) contacting the treated product solution with the fibrous Resin, and also comprising: an element for reducing the concentration of small organic compounds in a blood product while substantially maintaining a desired biological activity of the blood product, said element comprising highly porous adsorbent particles, which adsorbent particles are immobilized by an inert matrix.

0172About the Pathogen inactivating compound also includes their reactive Ab can construction products from the material, eg. as a blood product, for example, before Transfusion can be reduced.

0173the herein materials and elements as described can be used in Apheresemethoden will. Whole blood can in two or more specific components be separated (eg. as red blood cells, plasma and platelets). The term "apheresis" refers broadly to on procedures in which blood from a donor and is split into several components, the component (s) collected and recycled to keep interest is / are and the other components returned to the donor will. The donor receives replacement fluids while the reinfusion process to the compensation of the volume and pressure loss, which is caused by the removal of the components to support. apheresis are described in PCT Publication WO 96/40857 described.

Small organic compounds

0174On different-like assortment of small organic compounds adsorbed by the element of the present invention. The molecules may be cyclic his or acyclic. In one embodiment, the compounds preferably cyclic compounds such as psoralens, acridines or Dyes. In another embodiment, the compounds Thiols.

0175Non-limiting examples the cyclic compounds include actinomycin, Anthracyclinones, Mitomyacin, anthramycin, and organic dyes and photoreactive Compounds such Benzodipyrone, fluorenes, fluorenones, Furocumarine, Porphyrins, protoporphyrins, purpurins, phthalocyanines, Monostral Fast Blue, Norphillin A, phenanthridines, Phenazathioniumsalze, phenazine, Phenothiazines, phenylazides, quinolines and Thiaxanthenone. Preferably the compounds Furocumarine or organic dyes. preferred the compounds Furocumarine.

0176Non-limiting examples the Furocumarine include psoralens and psoralen derivatives. Specifically contemplated are 4'-aminomethyl-4,5 ', 8-trimethylpsoralen, 8-methoxypsoralen, halogenated psoralens Isopsoralene and quaternary amines, Sugar or other nucleic acid binding knotted groups Psoralens. are also considered the following psoralens: 5'-bromomethyl-4,4 ', 8-trimethylpsoralen, 4'-Bromomethyl-4,5 ', 8-trimethylpsoralen, 4 '- (4-amino-2-aza) butyl-4,5', 8-trimethylpsoralen, 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, 4 '- (2-aminoethyl) -4,5', 8-trimethylpsoralen, 4 '- (5-amino-2-oxa) pentyl-4,5', 8-trimethylpsoralen, 4 '- (5-amino-2-aza) pentyl-4,5', 8-trimethylpsoralen, 4 '- (6-amino-2-aza) hexyl-4,5', 8-trimethylpsoralen, 4 '- (7-amino-2,5-oxa) heptyl-4,5', 8-trimethylpsoralen, 4 '- (12-amino-8-axa-2,5-dioxa) dodecyl-4,5', 8-trimethylpsoralen, 4 '- (13-amino-2-aza-6,11-dioxa) -tridecyl-4,5', 8-trimethylpsoralen, 4 '- (7-amino-2-axa) heptyl-4,5', 8 -trimethylpsoralen, 4 '- (7-amino-2-axa-5-oxa) heptyl-4,5', 8-trimethylpsoralen, 4 '- (9-amino-2,6-diaza) nonyl-4,5', 8- trimethyl psoralen, 4 '- (8-amino-5-aza-2-oxa) octyl-4,5', 8-trimethylpsoralen, 4 '- (9-amino-5-aza-2-oxa) nonyl-4,5', 8-trimethyl psoralen, 4 '- (14-amino-2,6,11-triaza) tetradecyl-4,5', 8-trimethylpsoralen, 5 '- (4-amino-2-aza) butyl-4,4', 8-trimethylpsoralen, 5 '- (6-amino-2-aza) hexyl-4,4', 8-trimethylpsoralen and 5 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen. Preferably, the psoralen 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen.

acridines

0177Non-limiting examples the acridines include acridine orange, acriflavine, quinacrine, N1, N1-bis (2-hydroxyethyl) -N4- (6-chloro-2-methoxy-9-acridinyl) -1,4-pentanediamine, 9- (3-hydroxypropyl) aminoacridine, N- (9-acridinyl) glycine, S- (9-acridinyl) -glutathion. In a preferred embodiment is the acridine N- (9-acridinyl) -β-alanine, alternatively referred to as 5- (β-carboxyethyl) amino] acridine.

dyes

0178To non-limiting Examples include the dyes Phenothiazines such as methylene blue, neutral red, toluidine blue, crystal violet and Azure A, and Phenothiazone as methyl violet Bernthsen. Preferably the dye methylene blue or toluidine blue. More preferably, the Methylene blue dye.

Other organic compounds

0179the Concentration of a variety of organic compounds can be reduced will. Among other examples of compounds deleted Compounds. Methods for deleting undesirable Side reactions of pathogen inactivating compounds which contain a functional group that is an electrophilic group or is capable of forming such, are described in concurrently filed US patent application "Methods for Quenching Pathogen Inactivators in Biological Systems ", registration number 282173000600, filed on January 6, 1998. In this method, a material z. B. a blood product with a pathogen inactivating compound and a extinguisher treated, wherein the quencher comprises a nucleophilic functional group for covalent Reacting with the electrophilic group is capable of. In one embodiment, contains the pathogen inactivating Connecting a nucleic acid binding Ligand and a functional group, such as a mustard group, which is capable of reacting in situ to form the electrophilic group. Examples of the quencher count limited without it to be compounds including nucleophilic groups. Examples of nucleophilic groups include thiol, thioacid, dithioic, Thiocarbamate, dithiocarbamate, amine, phosphate and thiophosphate. The extinguisher , a nitrogen heterocycle be or include those such as. pyridine. The extinguisher may be a phosphate-containing compound, such as. for example, glucose-6-phosphate. The extinguisher may also be a thiol containing compound, including, but not limited on glutathione, cysteine, N-acetylcysteine, mercaptoethanol, dimercaprol, Mercaptan, mercaptoethanesulfonic and their salts, for. example, MESNA, homocysteine, aminoethane thiol, dimethylaminoethanethiol, Dithiothreitol and other thiol-containing compounds. Examples of aromatic thiol compounds include 2-Mercaptobenzimidazolsulfonsäure, 2-Mercaptonikotinsäure, Naphthalenthiol, Quinolinethiol, 4-nitrothiophenol and thiophenol. Other extinguishers nitrobenzylpyridine and inorganic nucleophiles such as Selenidsalze or Organoselenide, Thiosulfate, sulfite, sulfide, thiophosphate, pyrophosphate, hydrosulfide and Dithionitrit. The extinguisher may be a peptide compound containing a nucleophilic group. To the Example, the quencher be a cysteine ​​containing compound, for. example, a dipeptide such as about GlyCys, or a tripeptide, such as glutathione.

0180To other organic compounds can thiols count how as methyl thioglycolate, thiolactic acid, thiophenol, 2-mercaptopyridine, 3-mercapto-2-butanol, 2-mercaptobenzothiazole, thiosalicylic and thioctic acid.

EXAMPLES

0181the following examples serve to illustrate certain preferred Embodiments and Aspects of the present invention and are not of their way of limitation be understood framework.

0182In the following experimental description are the following Abbreviations: eq (equivalents); M (Molar); uM (Micromolar); N (normal); mol (moles); mmol (millimoles); .mu.mol (micromolar) nmol (nanomoles); g (grams); mg (milligrams); ug (micrograms); kg (kilograms); l (liters); ml (milliliters); ul (Microliters); cm (centimeter); mm (millimeters); (microns); nm (nanometers); min (minutes); . S and sec (seconds); J (Joules, also watt second); ° C (degrees Centigrade); TLC (thin layer chromatography); HPLC (high performance liquid chromatography); pHEMA and p (HEMA) (poly [2-hydroxyethyl]); PC (s) (platelet concentrate (s)); PT (prothrombin time); aPTT (activated partial thromboplastin time); TT (thrombin time); HSR (hypotonic shock response); FDA (United States Food and Drug Administration); GMP (good manufacturing practices); DMF (Drug Mastertiles); SPE (solid phase extraction); Aldrich (Milwaukee, WI); Asahi (Asahi Medical Co., Ltd., Tokyo, Japan); Baker (JT Baker, Inc., Phillipsburg, NJ); Barnstead (Barnstead / Thermolyne Corp., Dubuque, IA); Becton Dickinson (Becton Dickinson Microbiology system; Cockeysville, MD); Bio-Rad (Bio-Rad Laboratories, Hercules, CA); Cerus (Cerus Corporation; Concord, CA); Chrono-Log (Chrono-Log Corp .; Havertown, PA); Ciba-Corning (Ciba-Corning Diagnostics Corp .; Oberlin, OH); Consolidated Plastics (Consolidated Plastics Co., Twinsburg, OH); Dow (Dow Chemical CO .; Midland, MI); Eppendorf (Eppendorf, North America Inc .; Madison, WI); Gelman (Gelman Sciences, Ann Arbor, MI); Grace Davison (WR Grace &amp; Co., Baltimore, MD); Helmer (Helmer Labs, Noblesville, IN.); Maximum Celanese (Hoechst Celanese Corp., Charlotte, NC); internationally Processing Corp. (Winchester, KY); Millipore (Milford, MA); NIS (Nicolet, a Thermo Spectra Co., San Diego, CA); Poretics (Livermore, CA); Purolite (Bala Cynwyd, PA); Rohm and Haas (Chauny, France); Saati (Stamford, CT); Scientific Polymer Products (Ontario, NY); Sigma (Sigma Chemical Company, St. Louis, MO); Spectrum (Spectrum Chemical Mfg. Corp., Gardenia, CA); Sterigenics (Corona, CA); Tetko, Inc. (Depew, NY); TosoHaas (TosoHaas, Montgomeryville, PA); Wallac (Wallac Inc., Gaithersburg, MD); West Vacco (Covington, VA); YMC (YMC Inc., Wilmington, NC); DVB (divinylbenzene), LAL (Limulus Amoebocyte Lystate); USP (America American Pharmacopeia); EAA (ethylacetoacetate); EtOH (ethanol); HOAc (acetic acid); W (watts); mW (milliwatts); NMR (Nuclear Magnetic Resonance; spectra obtained at room temperature on a Varian Gemini 200 MHz Fourier Transform Spectrometer); ft³/ Min (cubic foot per minute); . Mp (melting point); g / min and gpm (gallons per Minute); UV (ultraviolet light); THF (tetrahydrofuran); DMEM (Dulbecco's Modified Eagle Medium); FBS (fetal Bovine serum); LB (Luria broth); EDTA (ethylenediaminetetraacetic acid); phorbol myristate acetate (PMA); Phosphate-buffered saline (PBS); AAMI (Association for the Advancement of Medical Instruments); ISO (International Standards Organization); EU (endotoxin units); LVI (large volume Syringes); GC (gas chromatography); M (mega); kGy (1000 Gray = 0.1 Mrad); Milliohms (MOhm); PAS III (platelet additive solution III); dH₂O (distilled water); IAD (immobilized Adsorption); SCD (sterile connection (connection) element).

0183One the examples below refers to HEPES buffer. This containing buffer 8.0 g of 137 mM NaCl, 0.2 g of 2.7 mM KCl, 0.203 g of 1 mM MgCl₂ (6H₂O), 1.0 g of 5.6 mM glucose, 1.0 g to 1 mg / ml bovine serum albumin (BSA) (available from Sigma, St. Louis, MO) and 4.8 g of 20 mM HEPES (available from Sigma, St. Louis, MO).

EXAMPLE 1

The fibrous resin Amberlite^® XAD-16

0184In be this example, both the kinetics of removal of aminopsoralens platelet and the platelet function and morphology using fibrous resin and elements, contain the non-immobilized Adsorbenskügelchen compared. Specifically was said immobilized Amberlite^® XAD-16 comprehensive fibrous Resin compared with elements that free (ie non-immobilized) Amberlite^® XAD-16 HP and Dowex^® XUS-43493 contained.

Preparation of filamentary resin and Adsorbenskügelchen

0185Maximum Celanese presented fibrous Resin obtained from Rohm and Haas Amberlite^® XAD-16 containing forth in a cleaned and hydrated state. the Fibers of the fiber network by Hoechst Celanese consisted of a Polyethylenterephthalatkern, and a nylon sleeve, wherein the sleeve has a lower melting temperature than the core had. The fibrous resin was prepared by adding tonearest the Adsorbenskügelchen uniformly in the fiber network were distributed. Next the fiber network was heated quickly, the polymer sleeve Fibers to melt and fuse to the Adsorbenskügelchen and other fibers is brought, which is a crosslinked fiber network revealed. The fibrous formed Resin contained the Amberlite^® XAD-16 at a loading of 130 g / m² (Ie each square meter of Fiber contained 130 g of Adsorbenskügelchen).

0186The fibrous Resin was cut into squares (14 cm × 14 cm), the resulting Portions of about 2.5 g of dry Amberlite®^® XD-16 contained. The Amberlite^® XAD-16 beads were then removed by immersion of the fibrous resin in 30% ethanol for about prewetted 10 minutes. The residual ethanol was then twice do the washing up with brine 10 Minutes away. Alternative methods of wetting the Amberlite^® XAD-16 and other adsorbents are also effective and are for the present Invention considered. It should be noted, that fibrous Resin, the other types of beads contains (Z. B. bridged or hyper crosslinked resins such as Dowex^® XUS-43493) no wetting step for a effective psoralen removal require.

0187Amberlite^® XAD-16 HP (High Purity) beads were also directly from Rohm and Haas in a cleaned and hydrated state based. For the loose (ie not immobilized) Amberlite^® XAD-16 HP-beads no pre-wetting before recording in a mesh bag was required; However, the mass of adsorbent was to take account of the water content the beads corrected (2.5 g dry = 6.8 g with 62.8% moisture). The Dowex^® XUS-43943 beads were purchased from Dow, the dry beads did not require wetting nor the mass of beads for water had to be revised. Polyester mesh bag (square of 7 cm × 7 cm; 30 micron openings) then were treated with 2.5 g (dry weight) either loose Amberlite^® XAD-16 HP or Dowex^® XUS-43493 beads filled.

0188The fibrous Resin and the adsorbent-containing pouches were by autoclaving in "wet" -Cyclus for 45 minutes at 121 ° C sterilized. Thereafter, the fibrous resin and the adsorbent-containing were Bag in separate, sterile, 1-liter PL 2410 Plastic container (Baxter) brought in. On the filling toward the PL 2410 Plastic containers were on a cross-flow bench using sterile scissors, hemostats, and an impulse sealer sealed.

Contacting the fibrous resin and Adsorbenskügelchen psoralen-containing Platelet concentrate (PC)

0189pools of platelet concentrate were prepared by combining 2-3 Units of single donor apheresis platelets in 35% autologous plasma / 65% Platelet additive solution (d. h. synthetic medium) prepared. This solution was the Aminopsoralen 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen added in an amount, with a final concentration of 150 uM 4 '(- 4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen was obtained. The resulting PC solution was divided into 300 mL units, and the units were then placed in PL 2410 Plastic containers (Baxter) introduced with 3 J / cm² irradiated at UVA. Following the irradiation, the treated PCs were in the PL 2410 Plastic container, either fibrous Resin with immobilized Amberlite®^® XAD-16, Wi Amberlite^® XAD-16 HP or Wi Dowex^® XUS-43493 contained, or into an empty PL 2410 Plastic container as transmit a control. The PL 2410 Plastic container (Baxter) was then placed on a Helmer-Blutplättcheninkubator at 22 ° C and stirred at about 70 cycles / minute.

0190the Samples of each PC were in 1 hour intervals during the first 8 hours of storage for analysis by HPLC of the remaining 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen removed. Each sample of PC was 5-fold with sample diluent dilute (Final concentration = 35% methanol, 25 mM KH₂PO₄, PH = 3.5) containing trimethylpsoralen (TMP) as the internal standard. Proteins and other macromolecules were by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were then centrifuged and the supernatant filtered (0.2 microns) and a C-18 reverse phase column (YMC ODS-AM 4.6 mm x 250 mm) by running a linear gradient of 65% solvent A (25 mM KH₂PO₄, pH = 3.5), 35% B (methanol) to 80% B analyzed in 20 minutes.

0191the Platelet yield while a 5-day Storage time with the fibrous Resin or a loose beads was daily by counting platelet on a Baker System 9118 CP (Baker Instrument Co .; Allentown, PA) monitored. Blood gases and pH were measured using a Ciba-Corning 238 pH / blood gas analyzer evaluated. The in vitro platelet function after 5 days of contact with the fibrous resin or the element, Free adsorbents contained, was determined by assay on the morphology, Shape change, hypotonic shock response, aggregation and GMP-140 (P-selectin) expression evaluated. The change in shape, Aggregation and hypotonic shock response were using a Lumi aggregometer (Chrono-Log) evaluated during the GMP-140 by flow cytometry using a Becton-Dickinson FACScan Fluorescence Analyzer (Becton Dickinson) was determined.

Psoralen removal and Platelet yield and function

0192In <figref idrefs="S121">5</figref> will the adsorption kinetics for 8-trimethylpsoralen, - the removal of 4 '(4-amino-2-oxa) butyl-4,5' of platelets synethetischem in 35% plasma / 65% medium (PAS III) with XUS-43493, XAD-16 HP and fibrous Resin containing XAD-16, are compared. Specific questions by the circles connected by the solid line indicated, Data represents the element, the non-immobilized Adsorbent XUS-43493 contained (2.5 g beads; &lt;5% moisture); represented by the triangles, connected by the dashed line, indicated data are for the Element, the non-immobilized XAD-16 HP acid (6.8 g beads; 62.8% moisture); and by, connected by the squares the dashed line indicated data are for the fibrous resin (Hoechst fibers with XAD-16 beads wetted in 30% ethanol; 14 cm × 14 cm). As the data in<figref idrefs="S121">5</figref> show the kinetics are of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen adsorption for both the element containing non-immobilized adsorbents, as also the element that the fibrous containing resin, very comparable. Therefore, the fiber formation process seems to be no significant influence on the removal kinetics to have.

0193It was also platelet yield and function of the fibrous Resin examined in comparison with the loose beads. Specific was at the Experimental th this study used: i) 6.8 g XAD-16 HP (62.8% moisture); ii) 14 cm × 14 cm fibrous XAD-16 (130 g resin / cm²), Moistened in 30% ethanol; and iii) 2.5 XUS-43493 (&lt;5% Moisture). Duplicate platelet units were for The XAD-16 HP and the fibrous Resin samples prepared, but only a single platelet unit for the XUS-43493-sample. The results are given in Table 1 below.

0194As shown in Table 1, the pH and pO₂-Values day 5 relative to the control on day 5 for samples containing non-immobilized globule (XAD-16 HP and XUS-43493) were slightly increased. The experiment with the fibrous Resin had pH and pO₂Values ​​on which the Control were comparable. Platelet counts showed a 9-22% platelet loss 5 days after contact with the fibrous medium and the element, the non-immobilized adsorbents contained. As shown in Table 1 specified, resulted in the fibrous Resin better yields (9% loss on the day 5), and revealed in all in vitro assays better performance compared to the element, the non-immobilized XAD-16 or XUS-43493-adsorbent contained.

table 1 <img img-content="tb" img-format="tif" he="65" wi="163" file="00530001.tif" />

0195Although the mechanism, the higher the pH and pO₂Values ​​underlying that observed for the element were the non-immobilized XUS-43493 and XAD-16 HP contained, does not have to be understood in order to practice the present invention to can, it is believed that the higher Values ​​by a slight decrease in the metabolism of blood platelets in The presence of the element that contains the non-immobilized adsorbent caused becomes. Compared were for The fibrous Media throughout pH and pO₂Values ​​on the day 5 obtained similar were to control than the XUS-43493- or XAD-16 beads.

0196the Platelet yields on day 5 were also for the fibrous Media relative to the XUS-43493- and XAD-16 HP-Adsorbenskügelchen better. The loss of 22-28%, the for the XUS-43493 media was observed was on several occasions observed. However, it should be noted that the presently preferred embodiment for a Element, the non-immobilized adsorbent with XUS-43493 contains the Transfer the platelet from this element after 8 hours Exposure includes, on which approach to loss platelets leads of &lt;5%.

0197the in Table 1 show data illustrated that the fibrous medium higher Platelet yield yields relatively to the elements, the non-immobilized adsorbents contain. Surprisingly results in the fibrous Medium also better platelet function on day 5, as shown by pH / pO₂, Change in shape, Aggregation, and GMP-morphology 140th Although an understanding of the basic principle for the increased Performance of the fibrous Medium for execution the present invention is not required, but can more Hypotheses are proposed. First, the fibers of which can surface the Adsorbenskügelchen are attached, an interaction between the platelets and the surface the beads hinder. Second, immobilization of the beads this prevent them from interacting and off mechanical effects, the for platelets are disadvantageous. Third, the immobilization of the beads a liquid shear on the surface the beads promote, whereby the interaction between the platelets and the surface of the globule is reduced; In comparison, non-immobilized beads with the liquid free-flowing, what to a small liquid stream relative to the surface the beads leads.

platelet loss

0198In view of the above data, it seems to be such that a certain variability in the platelet loss from one study to the next present. However, the on as a percentage of the control count Day 5 expressed platelet loss lower in studies in which the initial platelet count was higher. A study was carried out to show whether the number of platelets that is lost, for a given area of the material, to the platelet adhesion disposal stands is constant. For this Study, two platelet units pooled and divided the pool into two samples. A sample was half diluted with 35% autologous plasma / 65% synthetic media (PAS), so that the platelet count half the other unit was. The platelet mixture was treated with 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen + UVA treated and contacted with an element, the non-immobilized Adsorbent (2.5 g XUS-43493) for 5 days under the previously discussed standard platelet storage conditions contained.

0199the Total number of blood platelets, the lost, was for the two units nearly identical, whereas as a percentage calculated losses differed greatly. Thus, the results show that the platelet total loss to be constant for a certain time substantially seems; this means, while the percentage of platelet loss dependent on from the initial Platelet count varies, is the total number of platelet loss nearly be constant when equilibrium is reached. Based on specified in this example results showing the fibrous resin no negative effect on the in vitro platelet function.

EXAMPLE 2

The fibrous resin with activated carbon

0200In This example illustrates the kinetics of removal of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen of platelets and platelet function and morphology for fibrous Resin Amberlite^® XAD-16 comprises, and for fibrous resin, includes the immobilized activated carbon compared.

Preparation of the fibrous resin

0201Hochest Celanese prepared fibrous Resin Amberlite^® XAD-16 HP (Rohm and Haas) contained. The fibrous Resin Amberlite^® XAD-16 acid was as described in the previous example prepared, including the Wetting step with 30% ethanol. Hoechst Celanese prepared also a fibrous Resin immobilized activated carbon (Westvaco ((Luke, W. Va.)] At a loading of 375 g / m² (AQF-375-B) and 500 g / m² (AQF-500-B) contained. In a preferred embodiment, are the Adsorbent synthetic activated carbon, including,. B. Ambersorb and A-Supra. Synthetic activated carbons are due their F BILITY preferably, the amount of particulate Material that spread from the immobilized adsorption is eliminate. This fibrous resin was treated with a Method analogous to that for the fibrous Resin Amberlite^® XAD-16 contained prepared. The composition of the fibers for each fibrous Resin was the same.

0202The fibrous Resin was cut into squares (14 cm × 14 cm); the resulting Portions containing about 2, 5 g of dry Amberlite^® XAD-sixteenth Next was the fibrous Resin by autoclaving at "wet" cycles for 45 minutes at 121 ° C sterilized. Thereupon the fibrous resin in separate sterile 1-liter PL 2410 Plastic container (Baxter) introduced and the container on a cross-flow bench using sterile scissors, hemostats and an impulse sealer heat sealed.

Contacting the fibrous resin psoralen-containing PC

0203pools of platelet concentrate were prepared by combining 2-3 Units of single donor apheresis platelets in 35% autologous plasma / 65% synthetic medium (PAS III). 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen was added in an amount to provide a final concentration of 150 uM 4 '- butyl-4,5 (4-amino-2-oxa)', 8-trimethylpsoralen to achieve. The resulting PC solution was divided into 300 mL units, and the units were in PL 2410 Plastic container (Baxter) placed with 3 J / cm² irradiated at UVA. Following the irradiation, the treated PCs were in the PL transferred 2410 plastic container, the fibrous Resin with either XAD-16, AQF-375B, AQF 500B contained in or an empty PL 2410 Plastic container as a control. the PL 2410 Plastic containers were then on a Helmer-Blutplättcheninkubator at 22 ° C placed and stirred at about 70 cycles / minute.

0204As in the previous example, samples were each PC in 1-hour intervals while the first 8 hours of storage for analysis of residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralens means HPLC removed. Each sample of PC was 5-fold with sample diluent dilute (Final concentration = 35% methanol, 25 mM KH₂PO₄, PH = 3.5), the trimethylpsoralen (TMP) acid as the internal standard. Proteins and other macromolecules were by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were then centrifuged and the supernatant filtered (0.2 microns) and a C-18 reverse phase column (YMC ODS-AM 4.6 mm x 250 mm) by running a linear gradient of 65% solvent A (25 mM KH₂PO₄. pH = 3.5), 35% B (methanol) to 80% analyzed in 20 minutes.

0205the Platelet yield while a 5-day Storage time with the fibrous Resin or the element containing non-immobilized adsorbent was counting platelet monitored daily on a Baker System 9118 CP. Blood gases and pH were measured using a Ciba-Corning 238 pH / blood gas analysis element evaluated. The in vitro platelet function 5 days after contact with the fibrous resin or the element, the non-immobilized adsorbent contained, was determined by assay on the morphology, shape change, hypotonic shock response, aggregation and GMP-140 (P-selectin) expression evaluated. The change in shape, Aggregation and hypotonic shock response were using a Chrono-Log Lumi-aggregometer evaluated, whereas the GMP-140 by flow cytometry using a Becton-Dickinson FACScan Fluorescence Analyzer was determined.

Psoralen removal and Platelet yield and function

0206In <figref idrefs="S122">6</figref> will the adsorption kinetics for 8-trimethylpsoralen, - the removal of 4 '(4-amino-2-oxa) butyl-4,5' of blood cells synthetic in 35% plasma / 65% medium (PAS III) with fibrous resin XAD-16 contains, and fibrous Resin with two different loadings compared to activated carbon. Specific the data are given by the circles for the 4 '- (4-amino-2-oxa) butyl-4,5', 8- trimethyl psoralen removal with fibrous XAD-16 beads; the data given by the squares show the distance with fibrous AQF-500-B; and the data given by the triangles indicate the distance with fibrous AQF-375-B. As the data in<figref idrefs="S122">6</figref> show, are the kinetics of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl-psoralen adsorption for the various fibrous Resins very similar, and the fibrous resins showed all very good kinetics of removal (≤ 0.5 uM residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen after 4 hours).

0207the Platelet yield and in vitro platelet function for each the fibrous Resins was evaluated, and the data were summarized in Table 2 below.

table 2 <img img-content="tb" img-format="tif" he="73" wi="163" file="00580001.tif" />

0208Referring in Table 2 gave the coal-based fibrous resins good platelet yields in case of loss of less than 10%; as in the previous studies of the Example had the fibrous Resin containing XAD-16, a slightly higher platelet loss (about 17%). In terms of pO₂ are the day-5 values ​​for the on Coal based fibrous comparable resin control. Although not have to be understood, why the coal-based fibrous resin slightly higher pH values had to perform the present invention can, this can be a by residual extractables (eg., phosphate) act caused from the activation process unwanted pseudo result; the rapid rise of the pH value (pH = 7.3-7.4) of the following an 8-hour storage of the PC was observed with the coal-based fibrous resin, supports this idea. Experiments with USP coal extractable with less Substances are linked may the observed initial Increase of the pH eliminate.

0209The coal-based fibrous Resin gave good results both in the shape change and aggregation assays. Although the result for the change in shape the AGF-500-B-fibrous Resin is better than that of the control, with AQF-500-B showed associated platelets a slightly worse performance in the aggregation assay.

EXAMPLE 3

Effect of pHEMA coating to the adsorbent hemocompatibility

0210In '- (4-amino-2oxa) butyl-4,5' this example, the Entfernunskinetiken 4 are, 8-trimethylpsoralen of platelets and platelet function and morphology for both Dowex^® XUS-43493 and fibrous Resin Amberlite^® XAD-16, coated with pHEMA containing, compared.

Preparation of pHEMA coated Adsorbenskügelchen and fibrous resin

0211Dowex^® XUS-43493 (Known commercially as Optipore^® L493) containing about 50 wt .-% water, was obtained from Dow, and polymerized HEMA having a viscosity-average molecular weight of 300 kD was obtained from Scientific Polymer Products. Before the Coating were the Absorbenskügelchen to a water content dried of &lt;5%. A starting solution of pHEMA was prepared by dissolving of the polymer in 95% denatured ethanol / 5% water to give a pHEMA concentration prepared from 50 mg / ml.

0212Of the Coating process was International Processing Corp. in a 9-inch Wurster Flüssigbettbeschichter made with a load of about 4 kg (dry) of adsorbent. The coating process involved a pHEMA flow rate of 60-70 g / min, an inlet temperature of 50 ° C and an air flow rate of 200 ft³/ Min. Samples (50 g) of the coated adsorbent were during the Beschichtungspro zesses removed so that coating amounts The range of 3-18% (W / w) pHEMA were achieved; with 3.7%, 7.3% and 10.9% pHEMA (w / w) coated Adsorbenskügelchen were in the following bewritten studies used.

0213On Element, the non-immobilized dry (uncoated) of Dowex^® XUS-43493 (2.5 g) and pHEMA-coated Dowex^® XUS-43493 (3.0 g or 5.0 g) was by placing the desired Mass of adsorbent in a square 30 micron polyester mesh bag (7 cm x 7 cm) prepared. The adsorbent-filled Bags were in separate sterile 1-liter PL 2410 Plastic container (Baxter) introduced and heat-sealed with an impulse sealer. Thereupon, the adsorbent-filled bags, in the PL 2410 Plastic container were included, either by e-beam (NIS) or gamma irradiation (SteriGenics) sterilized at 2.5 Mrad; As indicated above, is the E-beam sterilization generally preferred.

0214Maximum Celanese prepared fibrous Resin Amberlite^® XAD-16 contained, according to the in Example 1 described method. The fibrous resin was cut into squares cut (14 cm × 14 cm); the resulting sections contained approximately 2.5 g of dry Amberlite^® XAD-sixteenth The Amberlite^® XAD-16 of the fibrous Resin was wetted simultaneously and coated with pHEMA by Immersion in a solution, the 50 mg / mL pHEMA / 5% distilled water contained in 95% ethanol. The remaining ethanol was rinsing twice with brine for 10 minutes away. This procedure resulted in a coating of about 6% (w / w) pHEMA. The fibrous Resin was then sterilized by autoclaving at "Moist" cycle for 45 minutes at 121 ° C. Thereafter, the fibrous was Resin in separate sterile 1-liter PL 2410 Plastic container (Baxter) introduced and on a cross-flow bench using sterile Scissors, hemostats and a pulse Siegler heat sealed.

Contact by pHEMA-coated Adsorbenskügelchen and fibrous Resin with psoralen-containing PC

0215pools of platelet concentrate were prepared by combining units of single donor apheresis platelets in 35% autologous plasma / 65% synthetic media (PAS III) prepared. 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen was added in an amount, with a final concentration of 150 uM 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen was achieved. The resulting PC solution was dissolved in 300 ml units split, and the units were in PL 2410 Plastic container (Baxter) placed with 3 J / cm² irradiated at UVA. Following the irradiation, the treated PCs were in the transmitted PL 2410 Kunststoffbehäter, either the non-immobilized pHEMA-coated Dowex®^® XUS-43493 containing element pHEMA coated fibrous resin with Amberlite^® XAD-16 contained, or in an empty PL 2410 Plastic container transmit a control. The PL 2410 Plastic container were then placed on a Helmer-Blutplättcheninkubator at 22 ° C and stirred at about 70 cycles / minute.

0216rehearse each PC were in 1 hour intervals during the first 8 hours of storage for analysis of residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen removed by HPLC. Each sample of PC was 5-fold with sample diluent dilute (Final concentration = 35% methanol, 25 mM KH₂PO₄, PH = 3.5) containing trimethylpsoralen (TMP) as the internal standard. Proteins and other macromolecules were by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were then centrifuged, and the supernatant was filtered (0.2 .mu.m) and a C-18 reverse phase column (YMC ODS-AM 4.6 mm x 250 mm) by running a linear gradient of 65% solvent A (25 mM KH₂PO₄. pH = 3.5), 35% B (methanol) to 80% analyzed in 20 minutes.

0217the Platelet yield after a 5-day Storage time with the fibrous Resin or the element containing non-immobilized adsorbent was by counting platelet determined on a Baker System 9118 CP. Blood gases and pH-value were evaluated using a Ciba-Corning 238 pH / blood gas analyzer. The in vitro platelet function after 5 days of contact with the fibrous resin or the element, the non-immobilized adsorbent contained, was determined by assay on the morphology, shape change, hypotonic shock response, aggregation and GMP-140 (P-selectin) expression evaluated. The change in shape, Aggregation and hypotonic shock response were using a Chrono-Log Lumi-aggregometer evaluated, whereas the GMP-140 by flow cytometry using a Becton-Dickinson FACScan Fluorescence Analyzer was determined.

Effect of pHEMA coating the psoralen removal

0218In <figref idrefs="S123">7</figref> will the adsorption kinetics for 8-trimethylpsoralen, - the removal of 4 '(4-amino-2-oxa) butyl-4,5' of platelets synthetic in 35% plasma / 65% medium (PAS III) with pHEMA coated and uncoated Dowex^® XUS-43493 beads compared. Specific is the 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen removal with 3.0 g Dowex^® XUS-43493, coated with 3.7% (w / w) pHEMA represented by the circles, being represented 7.3% (w / w) pHEMA by the triangles and 10.9% (W / w) pHEMA is represented by diamonds; are the squares for the 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen removal 2.5 g (dry) uncoated Dowex^® XUS-43,493th As the data in <figref idrefs="S123">7</figref> show, took the kinetics of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen adsorption with an increasing amount of pHEMA coating from. Although the mechanism for execution of the invention need not be understood, it is believed that this decrease to an increasing resistance to the Diffusion of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen caused in the interior of the adsorbent particles is.

0219the Adsorption kinetics of 4 '- (4-amino-2oxa) butyl-4,5', 8-trimethyl psoralen removal have also been with 5.0 g Dowex^® XUS-43493, coated with 3.7%, 7.3% and 10.9% (w / w) pHEMA determined. The results (Not shown) showed that the removal kinetics for those with 10.9% pHEMA coated beads to the uncoated (control) beads were comparable.

Effect of pHEMA coating on platelet yield

0220the Effect of pHEMA on platelet yield was in two studies using different amounts of adsorbent (3.0 and 5.0 g), but for the same amounts of pHEMA coating (3.7%, 7.3% and 10.9% [w / w]) determined. Platelet yields were relative to the platelet count on Day 5 for the treated PC that is not a non-immobilized adsorbent had been contacted through contained element calculated. As the gen zei results in Table 3, was when 3.0g tested on XUS-43493 were no nominal dose response at day 5 platelet yield with rising before pHEMA coating amounts; these results suggest that a small amount of the pHEMA coating most effective can be, since they have little effect on the 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen removal kinetics with preserved inhibition of platelet adhesion indicates the adsorbent surface. Unlike established with 3.0 g XUS-43493 nominal effect was a dose response was observed when 5.0 g were tested - rising pHEMA coating amounts increased the day 5 platelet yield. However, the yields are still lower than those in Using 3.0 g Adsorbenskügelchen were observed.

table 3 <img img-content="tb" img-format="tif" he="63" wi="166" file="00630001.tif" />

0221the suggest in Table 3 given results that the use a lower adsorbent (z. B. 2.5-3.0 g) in conjunction with a small amount of pHEMA coating (z. B. ≤ 3.0% w / w) is the best platelet yield will provide. As previously indicated, is the optimum amount of pHEMA coating equal to the minimum coating at which a Protective effect on platelet yield and in vitro platelet function is observed.

Effect of pHEMA coating and sterilization on platelet function

0222It has previously indicated that the sterilization methods a considerable can have an effect on the Adsorbensfunktion since the pHEMA coating crosslinked or cleaved who by irradiationthe can. Around evaluate impact, a study was carried out with elements, the non-immobilized pHEMA-coated (3.7% w / w) and uncoated XUS-43493, sterilized either 2.5 Mrad e-beam or 2.5 Mrad gamma-irradiation, contained. Each element contained 2.5 g (dry) of non-immobilized coated or uncoated XUS-43493 was added to a square 30 micron polyester mesh bag (7 cm × 7 cm); control included only treated PC, introduced in a PL 2410 plastic container. The results are summarized in Table 4 below.

table 4 <img img-content="tb" img-format="tif" he="83" wi="165" file="00640001.tif" />

0223Referring Table 4 appear on the pH values ​​on day 5 compared to the control to be very stable. The one with the element, the non-immobilized contains uncoated adsorbent measured pO₂Value is slightly increased, to a small close decrease in metabolism leaves; a coating with pHEMA seemed to reduce this effect, wherein the results for the sterilized with e-beam element, the non-immobilized Adsorbent contained, closer at the control lay than that for the gamma-sterilized item.

0224the platelet yields were for both pHEMA coated samples very well, the E-beam sterilized sample showed a slightly better performance. shape change and aggregation showed a pattern similar to that in the yield, wherein the element, the non-immobilized uncoated adsorbent contained, the lowest values ​​returned and the pHEMA-coated / e-beam sterilized Sample higher values ​​similar to the Control afforded. With an element, the non-immobilized Adsorbents contained, treated samples showed an equally good or better performance than the control in hypotonic Schockreaktions- (HSR) assay. Samples which were treated with the elements, the non-immobilized containing adsorbents gave lower morphology scores than the control, but showed lower activation levels as indicated by the GMP-140 assay.

0225A more Hämokompatibilitäts study was made in the elements, the non-immobilized uncoated XUS-43493 (2.5 g beads; 30 micron polyester mesh bag; 7 cm × 7 cm), uncoated fibrous Resin (14 cm × 14 cm), the Amberlite^® XAD-16 contained, and fibrous Resin pHEMA coated Amberlite^® XAD-16 contained, were compared. The favorable effect of pHEMA on the fibrous Resin is indicated by the results shown in Table 5 below.

table 5 <img img-content="tb" img-format="tif" he="82" wi="165" file="00660001.tif" />

0226As regarding the data the in vitro platelet function and the platelet yield shown in Table 5, the coating with pHEMA brought the pO₂values ​​for the fibrous Harz closer to the for control observed zoom. The pHEMA-coated fibrous resin also showed a higher platelet yield as the uncoated fibrous Resin. The results show that the pHEMA-coated fibrous resin better performance provided than the uncoated XUS-43493 beads in all the assays of the in vitro platelet function. About that , the uncoated fibrous resin yielded better performance than the uncoated XUS-43493 beads in most assays the in vitro platelet function.

EXAMPLE 4

Effect of glycerol and Polyethylene glycol on the adsorption capacity

0227In This example illustrates the effect of glycerol and polyethylene glycol as stabilizers in the adsorption capacity and the kinetics of removal examined by aminopsoralens from plasma. Free (ie non-fibrous) Amberlite^® XAD-16 and Dowex^® XUS-43493-Adsorbenskügelchen were used in the experiments of this example.

methodology

0228Amberlite^® XAD-16 HP (Rohm & Haas (Philadelphia, PA)) and Dowex^® XUS-43493 (Supelco, Bellefonte, PA) were to <5% Water in an oven at 80 ° C dried. Known masses of adsorbent were soaked in ethanol solutions 0-50% Glycerol, 50% PEG-200 or 50% PEG-400 (glycerol, PEG-200 and PEG-400 containing from Sigma). Following a 15-minute incubation period, at room temperature the excess solution was removed and the samples via Overnight in an oven at 80 ° C dried; drying of the adsorbent at temperatures> 120 ° C was avoided, as previously changes in the adsorbent properties (z. B. bad melting behavior) at higher Temperatures were observed. After drying, the Adsorbensproben were weighed to the mass of stabilizer per mass of adsorbent to determine.

0229Several Individual studies have been made. Control samples of "non-wetted" adsorbent and "optimal wetted" adsorbent were included in the study, as described below. at the non-wetted samples of adsorbent it was dried Adsorbent which had been subjected to no pre-treatment, whereas the optimal wetted samples of adsorbent by wetting the absorbent with 30% ethanol / 70% dH₂O prepared had been. The optimal Wetting adsorbent was with dH₂O to remove residual ethanol flushed. The adsorbent was prepared right in front of Adsorptionsuntersuchung, That no drying was carried out to make sure.

0230Each the adsorption studies was carried out using 100% human plasma made the 150 uM 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, added ³H-4 '- (4-amino-2-oxy) butyl-3,5', 8-trimethylpsoralen, contained. Plasma (6.0 ml) was placed in vials with various stabilizers treated adsorbent contained. the were Adsorbensmassen for corrects the glycerol or PEG content to 0.2 g of adsorbent to obtain. The vials were placed on a rotator and stirred at room temperature. Plasma samples were taken at various time points, and the amounts of residual ³H-4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen certainly. Samples (200 ul) were in 5.0 ml Opti Phase HiSafe Liquid Scintilla tion Cocktail diluted (Wallac) and on a Wallac 1409 liquid scintillation counter (Wallac) counted.

adsorption capacity of Amberlite^® XAD-16 and Dowex^® XUS-43493, treated with glycerol

0231In <figref idrefs="S124">8th</figref> becomes the effect of pretreatment with ethanol solutions containing various amounts contain of glycerol, on the relative adsorption capacity of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen in 100% plasma for Amberlite^® XAD-16 and Dowex^® XUS-43493 compared. The Adsorbensproben were wetted in ethanol / glycerol solutions for 15 minutes, then for 48 Hours at 80 ° C dried. Individual measurements of adsorptivity were made 4 hours after contact. Referring to<figref idrefs="S124">8th</figref>. are the x on the axis Glycerol content shown the weight / volume percent of glycerol in ethanol. The adsorption capacities shown on the y-axis are percentages relative to the adsorption capacity of optimally wet Adsorbensprobe. The adsorption of XUS-43493 is represented by the squares, whereas the is reproduced from XAD-16 by the circles.

0232As the data in <figref idrefs="S124">8th</figref> show is the power of XAD-16 from about 30% increases in the dry sample to over 90% in the sample, the in a 20% glycerol solution had been wetted. These results show that very small amounts of glycerol for maintaining a high adsorption capacity after are drying required. Control samples in 50% ethanol / 50% dH₂O (no glycerol) wetted before drying were, adsorption capacity showed similar to untreated samples that had been dried. In contrast to the XUS-43493 samples showed no effect of glycerol on the adsorption capacity; adsorptivity approaching to 100% of all amounts of glycerol. Although for the execution of the the present invention is not critical, this supports Monitoring the hypothesis that glycerol acts to a to prevent shrinkage of the adsorbent pores during drying; because XUS-43493 has a highly crosslinked structure, subject it during no drying shrinkage of the samples.

0233Rehearse, treated with glycerol, seemed very stable to to be drying. No changes adsorptivity were observed in samples for 7 days on a crossflow Bank been stored products (data not shown).

0234In a preferred embodiment, of the present invention, 2.5 g dry weight of adsorbent for the removal of psoralen and psoralen photoproducts from each unit platelets used. The soaking of the adsorbent in 30% glycerol / 70% ethanol, followed by drying, results in an adsorbent which about 50% glycerol contains. A sample of the adsorbent of 5.0 g would therefore 2.5 g dry adsorbent and 2.5 g glycerol contained. Therefore, a typical unit of platelets 300 ml 0.8% glycerol containing an amount for transfusion deemed acceptable will.

adsorption capacity of Amberlite^® XAD-16 and Dowex^® XUS-43493, treated with glycerol or PEG

0235additional Studies were performed with the low molecular weight polyethylene glycols PEG-200 and PEG-400 made which agents of low toxicity are, the non-volatile soluble in ethanol and water are. The adsorbent of the samples were 15 minutes in 50% solutions of treated PEG-400, PEG-200 or glycerol in ethanol. In<figref idrefs="S125">9</figref> becomes the effect of the stabilizers on the adsorption capacity of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen with dried adsorbent in 100% plasma for Amberlite^® XAD-16 (Below) and Dowex^® XUS-43493 (top) compared; the samples that were not wet, are marked with "no Tx". The adsorption is as a percentage relative to the capacity of optimally wet adsorbent reported.

0236As by the data in <figref idrefs="S125">9</figref> and indicated on the basis of his "macroreticular" structure predictably, was the capacity of Dowex^® XUS-49493 by drying does not affect ( "no Tx" sample). Conversely pointed Amberlite^® XAD-16 about 35% of the maximum capacity when it was dried. Treatment of XAD-16 with glycerol, PEG-200 and PEG-400 improved in all cases, the capacity of getrock Neten Absorbent; the Adsorbenskapazitäten with each of which were more than 90%, Glycerol> PEG-200> PEG-400th Although the precise Mechanism for execution the present invention does not have to be understood, there may be differences capacity between the glycerol and the two PEG solutions by decreasing Penetration of the stabilizer with increasing molecular weight be effected. This means, that while the 15-minute Application process, the glycerol (MW = 92.1) for more complete Penetrating the adsorbent pores may be capable as both PEG-200 (MW = 190-210) or PEG-400 (MW = 380-420) the slower due to their larger size diffuse.

Adsorgtionskinetiken of Amberlite^® XAD-16, treated with glycerol or PEG

0237A Investigation has also made to determine whether the filling of the pores of the adsorbent with glycerol or PEG results in reduced adsorption kinetics. In<figref idrefs="S126">10</figref> is the adsorption of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen via a 3 hours Period from 100% plasma using Amberlite®^® XAD-16 compared that had been wetted in a plurality of different solutions. Specifically, the data in <figref idrefs="S126">10</figref> for XAD-16 i) wet prior to drying with a 50% solution of glycerol (open squares, connected by solid lines), ii) wet prior to drying with a 50% solution of PEG-400 (black circles connected by dashed lines), iii) pre-wetted, ie directly in front of the investigation, with 50% ethanol / 50% dH₂O (black triangles, connected by dashes), and iv) no treatment (black Squares connected by solid lines; "No Tx"). The data in<figref idrefs="S126">10</figref> show, that Amberlite^® XAD-16 samples which wets in 50% glycerol / 50% ethanol or 50% PEG-400/50% ethanol solutions had been, had adsorption kinetics, which in very dense of the sample were, which had been wetted optimally in ethanol (ie the pre-wetted with ethanol sample). The XAD-16 sample dried but was not treated (no Tx) achieved after 3 hours a mere 30% removal.

0238the show in this example, data presented that treatment Amberlite^® XAD-16 with stabilizers in the form of solutions, containing 50% ethanol and 50% glycerol, PEG-200 or PEG-400 the prevent the drying-related loss of adsorptive can. Laying The results obtained with these stabilizers suggest that low molecular weight wetting agents useful methods deliver improving Adsorbensfunktion.

EXAMPLE 5

Removal of methylene blue FFP

0239This Example is directed to the ability of a variety of polymeric adsorbent, methylene blue remove from fresh frozen plasma.

0240the Experiments of this example evaluated "free" adsorbent resin (ie not in an element, containing non-immobilized adsorbents added) and fibrous resin out. The tested free adsorbent resins were Amberlite^® XAD-18 HP (Rohm and Haas), MN-200 (Purolite) and Dowex^® XUS-43493 (Dow Chemical Co.). The XAD-16 HP was in a hydrated state before, so that no pretreatment (ie, no wetting) is necessary was, and the MN-200 has also been provided in a fully hydrated state; the XUS-43493 was dry.

0241XAD-16 containing fibrous Resin was prepared as generally described in Example 1. Short a 2 cm × 7 was told cm (ie 14 cm²) Strips of fibrous resin, of 130 g / m² XAD-16 contained, initially in wetted 70% ethanol and then fully rinsed in distilled water.

0242A Ausangslösung of methylene blue (10 mM) was prepared by dissolving methylene blue USP (Spectrum) was prepared in distilled water. The starting solution of Methylene blue was added to a sample of 100% plasma to a final concentration 10 uM added. Samples of the "free" adsorbent resin (d. h. XAD-16 HP, MN-200 and XUS-43493) were dissolved in 50 ml polypropylene tubes for adsorption weighed. The water content of each adsorbent was determined by measuring the Mass loss determined during drying. The mass of each adsorbent was for corrects the water content, so that in each case the equivalent was used by 0.25 g Trockenabsorbens.

0243A 30-ml sample of 100% plasma, containing 10 uM methylene blue, was each vial was added. The vials were placed on a rotator at room temperature. Samples (200 ul) were taken from each vial at 15-minute intervals and assayed for residual methylene blue by HPLC. Each Sample of the plasma was fivefold with sample diluent dilute (Final concentration = 35% methanol, 25 mM KH₂PO₄, PH = 3.5). Proteins and other macromolecules were by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were centrifuged and the supernatant filtered (0.2 microns) and on a C-18 reverse phase column (YMC ODS-AM, 4.6 mm × 250 mm) by running a linear gradient of 65% solvent A (25 mM KH₂PO₄. pH = 3.5), 35% B (methanol) to 80% B in 20 minutes analyzed. The detection limit for the HPLC assay was about 0.5 uM Methylene blue.

0244In <figref idrefs="S127">11</figref> are the kinetics of adsorption of methylene blue over a 2-hour Period from 100% plasma compared. Referring to<figref idrefs="S127">11</figref> are the XAD-16 HP data represented by unfilled diamonds, which are connected by dashed lines, the MN-200 data by black triangles connected by solid lines are, the XUS-43493 data represented by open circles, by Dashed lines are connected and the fibrous resin, containing XAD-16, represented by black squares connected by solid lines are. As the data show, the XAD-16 HP and MN-200 data indicated the fastest adsorption kinetics, followed by XUS-43,493th the slightly slower kinetics of the XUS-43493 may a result of the slower be wetting, as it was used in the dry state. Finally pointed the fibrous Resin containing XAD-16 adsorption kinetics slowest on. This may be the result of poor contact between the fibrous Resin and the plasma during be the Chargeninkubation, since a part of 14 cm²-Streifens of the fibrous Resin is not completely in the plasma during the Adsorptionsuntersuchung was immersed what the effective contact area between the adsorbent and the plasma decreased.

0245the Data show that not psoralen concerned pathogen inactivating Compounds such as the phenothiazine dyes from blood products under Use of the resins of the present and for use in the Invention be removed in contemplated resin fibrous can.

EXAMPLE 6

Removal of acridine compounds of packed red cells

0246This Example is directed to the ability of a variety of resin materials, acridine compounds of packed red cells (PRBCs) to remove. More specifically evaluate the experiments this acridine [(β-carboxyethyl) amino] - example of the removal of the acridine compound 5 from PRBCs from.

0247the chemical structures of various acridines are in <figref idrefs="S128">12</figref> shown. As in<figref idrefs="S128">12</figref> stated 9-aminoacridine and 5 - [(β-carboxyethyl) amino] acridine Aminoacridines.

resin selectivity

0248the Equilibrium adsorption of the compound 5 - [(β-carboxyethyl) amino] acridine was studied with various types of resin. The ausgewerteren polymeric adsorption resins were Amberlite^® XAD-2, XAD-4, XAD-7 and XAD-16 HP (Rohm and Haas); Purolite^® MN-150, MN-170, MN-200, MN-300, MN-400, MN-500 and MB-600; and Dowex^® XUS-43493 and XUS-40285 (Dow Chemical Co.). In addition, various Amberlite were^®-Anionenaustauscherharze (IRA-958, IRA-900, IRA-35, IRA-410 and IRA-120, Rohm and Haas) and Amberlite^® weak Cation exchange resin (DP-1, Rohm and Haas) were tested. Furthermore more coals were evaluated, including Hemosorba^® AC (Asahi), PICA G277 and Norit A Supra (both commercially obtainable of American Norit). After all was also Porapak^® RDx (Waters), a Styrolvinylpyrrolidon copolymer, which has an affinity aromatic having nitro compounds tested.

0249was initially a Gleichgewichtsadsorptions study with samples of each resin made to the capacity acridine [(β-carboxyethyl) amino] - 5 and adenine (6-aminopurine) evaluated. About 0.1 g of resin was weighed and transferred to a 6 ml polypropylene tubes. A 5.0 ml aliquot of 25% plasma / 75% Adsol^® (Baxter), 100 uM acridine [(β-carboxyethyl) amino] - 5 of in distilled water, then each tube was added. Cell products such as red blood cells are typically stored in a medium containing a low percentage of plasma (10-35%) having an equilibrium amount contains on synthetic medium. Adsol^® is an example of a synthetic medium consisting of adenine, dextrose and mannitol in a saline solution consists. Naturally the present invention contemplates the use of other concentrations the acridines than 100 uM in mixtures with plasma and other synthetic media into consideration. Next The tubes were on a tumbling shaker placed and incubated for 3 hours at room temperature. After incubation, aliquots of each sample for analysis of remaining 5 - [(β-carboxyethyl) amino] acridine and adenine removed by HPLC.

0250For the HPLC method each sample was two-fold with sample diluent (50% methanol, 25 mM KH₂PO₄, pH = 3.5), washed and the proteins and other macromolecules were prepared by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were then centrifuged, and the supernatant was filtered (0.2 .mu.m) and a C-18 reverse phase column (YMC ODS-AM 4.6 mm x 250 mm) by running a linear gradient of 75% solvent A (25 mM KH₂PO₄. pH = 3.5), 25% B (methanol) to 80% B in 20 minutes analyzed. For the Removal of 5 - [(β-carboxyethyl) amino] acridine were estimated capacities (Mol / g) at C_r = 1 uM of each adsorption with C₀ = 100 uM determined; the results are in the second column of Table 6 is reproduced (NN = not detectable). For the Removal of adenine was the estimated capacity (mmol / g) at C_r = 1 mM of each adsorption measurements with C₀ = rated 1.5 mM. The results are in the third column of the table 6 reproduced (NN = not detectable).

table 6 <img img-content="tb" img-format="tif" he="133" wi="167" file="00750001.tif" />

0251indeed is contemplated the use of any resin for Adsorbing acridine compounds capable of adsorbing but preferred Resins selective 5 - [(β-carboxyethyl) amino] acridine opposite to Adenine and exhibit low hemolysis. In <figref idrefs="S129">13</figref> are the data for the Adeninkapazität (y-axis) and capacity of 5 - [(β-carboxyethyl) amino] acridine (X-axis) for various resins applied. As shown by the data in Table 6 and <figref idrefs="S129">13</figref> stated, reported the Dowex^® XUS-43493- and Purolite^® MN-200 resins which highest capacity acridine [(β-carboxyethyl) amino] - 5 on; were about addition, both high capacity of 5 - [(β-carboxyethyl) amino] acridine and pulled a small Adeninkapazität into consideration, showed Amberlite^® XAD-16 HP performed well.

0252the Results from those described in this example, in vitro experiments suggest that Dowex^® XUS-43493 and the related resin Purolite^® MN-200 preferred resins for represent the removal of acridine compounds from PRBCs.

EXAMPLE 7

Removal of acridine compounds of packed red cells with styrene-divinylbenzene adsorbents

0253This Example is directed to the ability of a variety of different Styroldivinylbenzol- (styrene-DVB) -Adsorbenzien, remove aminoacridine compounds from blood products. Specific evaluate the experiments of this example, the removal of acridine and 9-amino acridine (depicted in <figref idrefs="S128">12</figref>) from a plasma / Adsol^®-Solution out.

A. Experimental Procedure

0254For the experiments, of this example were the starting solutions (10 mM) of 5 - acridin [(β-carboxyethyl) amino] (Cerus) and acridine orange (Aldrich) were prepared in distilled water, and a starting solution (10 mM) of 9-aminoacridine (Aldrich) was prepared in ethanol. acridine [(β-carboxyethyl) amino] - The 5 was a solution was added, containing 25% plasma / 75% saline to a final concentration 100 uM to obtain, whereas the acridine orange and 9 amino acridine compounds solutions were added, the 25% plasma / 75% Adsol^®- (Baxter) -Red Blood cell preservative solution contained, an acridine final concentration to achieve 100 uM.

0255the Adsorbents used were Amberlite^® XAD-16 HP (Rohm and Haas); Purolite^® MN-200 and Dowex^® XUS-43493 (Supelco). The water content of each adsorbent was determined by measuring the Mass loss determined during drying; the water content was so corrected to the equivalent 0.25 g dry weight of each adsorbent was used. The adsorbents were dissolved in 50 ml Falcon tubes accurately weighed. Thirty (30) ml of 25% plasma / 75% Adsol^®Solution, the 100 uM acridine contained, each adsorbent-containing tubes were added. The tubes were then on a rotary device placed at room temperature and the solutions 500 ul samples were removed at various time points and stored for later analysis.

0256rehearse were analyzed by HPLC for the levels of residual acridine. Each sample was two-fold with sample diluent (50% methanol, 25 mM KH₂PO₄, pH = 3.5), washed and the proteins and other macromolecules were prepared by incubating the samples at 4 ° C. for 30 precipitated minutes. The samples were then centrifuged, and the supernatant was filtered (0.2 .mu.m) and a C-18 reverse phase column (YMC ODS-AM 4.6 mm x 250 mm) by running a linear gradient of 75 solvents A (25 mM KH₂PO₄. pH = 3.5), 25% B (methanol) to 80% B in 20 minutes analyzed. Detection was by visible absorbance using a detector sets with diode array at 400 nm for 9-amino acridine, 490 nm for Acridine and 410 nm for 5 - [(β-carboxyethyl) amino] acridine.

B. adsorption kinetics

0257the Kinetics of adsorption of 5 - [(β-carboxyethyl) amino] acridine over a 3-hour period of 25% plasma / 75% saline were for Dowex^® XUS-43493, Purolite^® MN-200 and Amberlite^® XAD-16 HP compared. In<figref idrefs="S130">14A</figref> and <figref idrefs="S131">14B</figref> is each of the remaining 5 - [(β-carboxyethyl) amino] acridine represented as a function of time, wherein <figref idrefs="S131">14B</figref> the data on a logarithmic scale shows. In<figref idrefs="S130">14A</figref> and <figref idrefs="S131">14B</figref> are The XAD-16 HP data shown by black circles, which through Dashed lines are connected, the MN-200 data by black shown squares connected by dashed lines, and the XUS-43493 data represented by black triangles are connected by solid lines. As the data show, revealed XUS-43493 and MN-200 the fastest adsorption kinetics and were almost equivalent. XAD-16 HP appears less capacity for 5 - acridine [(β-carboxyethyl) amino] exhibit.

0258In <figref idrefs="S132">15</figref> be the adsorption kinetics for removal of 9-amino acridine and acridine orange from 25% plasma / 75% Adsol^® With Dowex^® XUS-43493 compared. Referring to<figref idrefs="S132">15</figref> to stand the black squares with the dashed lines for 9-amino acridine and the black circles with solid lines for acridine. As the data show the amounts of 9-aminoacridine addition over 3 hours were not detectable. In comparison, the capacity of Dowex was^® XUS-43493 for Acridinorange lower, with respect to the presence of two tertiary amino groups can stand on Acridinorange.

0259the Results of this study have the general ability the polystyrene-DVB adsorbents after removing acridines from blood products. It did note be taken that Wechinteractions between the chemical agent (Z. B. the acridine) are determined and the cell-containing blood preparation, whether be a batch or flow-adsorption most effectively becomes. In a similar way be the Hämokompatibilitäts properties individual adsorbents affect the determination of which Adsorbents are most effective.

0260It should be understood that the invention is not exact to the Details of operation or exact compounds, compositions, Methods or procedures, be as shown and described, limited is, as modifications and equivalents one skilled in the art will be apparent. From the above it should be understood that the methods and elements in apheresis and other elements and procedures that are currently for processing of blood products for transfusion are used, can be added.

EXAMPLE 8

0261In This example illustrates the use of various types of powdered carbon compared as the active component in the medium, and shown that a Careful selection of the active ingredient for reducing the concentration a small organic compound and retention of biological function the liquid is required. The five Example 8 illustrated activated carbons reduce the concentration the psoralen, 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen, in plasma by about the same amount. Where "A Supra" is used as the adsorbent is, however, there is a much better retention of clotting factors relative to the other adsorbents.

02624 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen the plasma was added to a concentration of 150 uM, and the plasma was in 325 ml-batches with 3.0 J / cm² UVA irradiated to inactivate pathogens. The residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen concentration was about 90 uM in the resulting plasma pool. 325 ml of the irradiated plasma were at 20 ml / min through 5 different types of Cuno Zeta Plus 90 mm carbon pads pumped. The clotting factor levels in plasma, clotting times and 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl-psoralen concentration were examined before and after each flow through the KohlenstoffPads.

<img img-content="tb" img-format="tif" he="63" wi="165" file="00790001.tif" />

Charcoal specifications <img img-content="tb" img-format="tif" he="46" wi="167" file="00790002.tif" />

0263the Water flow rate R10S media is 2 gallons water / min / Foot² at a pressure differential 1.5 psi.

0264Remarks: The A Supra Quality was prepared with the same specifications as the R1xS series; only the carbon has been replaced. PTT is partial thromboplastin time, with an increase in a distance the coagulation factors indicating. I, VIII and IX to be certain coagulation factors. All values ​​are relative to the post-photochemical Treatment given. 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen was analyzed by HPLC, and the coagulation factors and times were examined by means of a coagulation analysis machines.

EXAMPLE 9

0265In this example, the use of two particle sizes is compared and the better performance of the smaller particles in reducing the concentration of a small organic compound and the tradeoff detected with the biological function of the liquid. A Particle size of less than 50 microns results in a significant increase of the percentage of from plasma remote psoralen relative to the use of a particle size of between 100 and 50 microns: 99.2% compared to 96.1%.

0266photochemical treated plasma was similarly prepared as in Example 8, Zeta-Plus-like Filters were prepared, the two different particle sizes of ground Dowex Optipore L493 instead of the powdered Activated carbon according to the Cuno R1xS specifications contained. 325 ml photo chemically treated Plasma was pumped through each pad at 20 mL / min. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="40" wi="163" file="00800001.tif" />

EXAMPLE 10

0267In This example describes the effect of a change in the mass fraction the active component in reducing the concentration of a small organic compound and the tradeoff with the biological function the liquid compared.

0268photochemical treated plasma was similarly prepared as in Example 8. FIG. A Supra pads were the standard R1xS loading of about 60% carbon and a lower loading level of 30% prepared. Approximately 230 ml of plasma was pumped through each 90 mm disc. The 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="40" wi="165" file="00810001.tif" />

EXAMPLE 11

0269This Example demonstrates that in some cases the treatment of the medium with a hydrophilic polymer benefits for the biological function can show. Photo Chemically treated plasma was similarlyin Example 8 prepared. A 90 mm-R14S pad was Night with a 50 mg / ml solution purged of polyhydroxyethyl methacrylate (pHEMA) in 95% ethanol, in a Oven at 70 ° C dried until no further weight change was more. The second pad was rinsed with 95% ethanol without corresponding pHEMA and dried. About 325 ml of plasma was pumped through the disks at 5 ml / min. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="35" wi="135" file="00810002.tif" />

EXAMPLE 12

0270In This example shows that the flow rate has an effect on the reduction of the concentration of the small organic compound can show.

0271photochemical treated plasma was similarly prepared as in Example 8. FIG. A Supra pads were the standard R1xS loading of about prepared 60% carbon, and 325 ml of the treated plasma were each of the disks 47 mm diameter pumped at three different flow rates. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="47" wi="160" file="00820001.tif" />

EXAMPLE 13

0272In This example shows how the volume of liquid has an effect both to the reduction in concentration of small organic can have connection as well as the biological activity of the liquid. The critical Nature of the form of contact between the biological fluid and the reduction element the organic molecule is illustrated. Shown is the effect that the liquid volume both to the reduction in the concentration of small organic Compound as well as the biological activity of the liquid shows. Where the by the element-pumped plasma volume is increased (eg. as doubles) is about the same concentration of small organic compounds, z. B. psoralen from the solution reduced, whereby a significant increase in the retention the coagulation factors occurs.

0273photochemical treated plasma was similarly prepared as in Example 8. FIG. Zeta-Plus-like Filters were with ground Dowex Optipore L493 with particles of less than 50 microns instead of the powdered Activated carbon according to the Cuno R1xS specifications prepared. The plasma was pumped through the filter at 20 ml / min and the Samples were taken at 180 ml and 325 ml. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="40" wi="165" file="00830001.tif" />

EXAMPLE 14

0274In this example, the use of a sintering medium will be described.

0275slices ninety millimeters in diameter by 1/4 inch thick were of Porex generated. The disks contained various weight fractions of finely ground Dowex Optipore L493 with particle sizes between 20 microns and 100 microns and small particles of ultrahigh molecular weight polyethylene (quality UF220) which were then sintered together. Photo Chemically treated plasma was similar prepared as in Example 8, and 200 ml of plasma was prepared by each of the disc at 16 - 18 ml / min pumped. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="47" wi="165" file="00830002.tif" />

EXAMPLE 15

0276In This example describes the effect of increasing the mass of adsorbent by increasing the diameter of the filter described with a constant thickness.

0277photochemical treated plasma was similarly prepared as in Example 8. FIG. Approximately 325 ml of the treated plasma were slices of 90 and 47 mm diameter of a R03S-quality pumped at 5 ml / minute. 8-trimethylpsoralen - '(4-amino-2-oxa) butyl-4,5', the 4 and clotting factors were analyzed as in Example 8. FIG.

<img img-content="tb" img-format="tif" he="30" wi="168" file="00840001.tif" />

EXAMPLE 16

Preparation of PRBCs treated with a 5 - [(β-carboxyethyl) amino] acridine derivative and glutathione

0278pools of PRBC were prepared from fresh ABO-adapted thoroughbred. The units of whole blood were at 3800 rpm for 5 minutes using a Beckman Sorvall centrifuged RC3B centrifuge. The plasma was measured using a Auspressgeräts in a another clean bag pressed. theUnits of PRBCs were in a Clintec Viaflex bag of 3.0 l size pooled, if more than one Unit was required. For each unit of PRBCs were 94 ml Erythrosol added. The percentage hematocrit (HCT) was filling a capillary tube with the blood sample and 5 minutes Centrifuging measured. hematocrit was determined to make sure that he did not fall below 55%. 3.3 ml were added at 12% of glucose per 100 ml of PRBCs. the final percent hematocrit were determined. PRBC (300 ml) was PL146 in plastic container (Baxter Healthcare) decanted. The blood bags were added to 6.0 ml of 150 mM glutathione to a final concentration of 3.0 mM and 3.0 ml of a 30 mM 5 - [(β-carboxyethyl) amino] acridine derivative for a Final concentration of 300 uM to reach. The PRBC mixture was 1 minute using a wrist shaker (manufacturer) shaken. The 5 - [(β-carboxyethyl) amino] acridine derivative and glutathione treated PRBCs were added at room temperature overnight incubated the 5 - [(β-carboxyethyl) amino] acridine derivative 5 - [(β-carboxyethyl) amino] acridine dismantle.

EXAMPLE 17

acridine [(β-carboxyethyl) amino] - HPLC assays for 5 in PRBC and PRBC supernatant

0279the Sample (100 ul) was charged with 100 .mu.l saline solution dilute and the resulting mixture vortexed. The solution was added 300 ul of 20 mM H₃PO₄ in CH₃CN was added and the mixture 15 seconds vortexed long. The sample was centrifuged at 13,200 rpm for 5 minutes. The supernatant (200 ul) was in 800 ul diluted in cold 0.1 M HCl and vortexed. The sample was in an autosampler vial Using a 0.2 micron Gelman Acrodisc filter filtered. The HPLC conditions were as follows: (.. model and part number for column and general column) Column = Zorbax SB-CN, 4.6 mm × 150 mm, 3.5 micron particles; guard column = (4.6 mm × 12.5 mm, 5 .mu.m Particles (Mac Mod Analytical, Inc. (Chadds Ford, PA)); the mobile phase for A was 10 mM H₃PO₄ in HPLC grade water; the mobile phase for C was 10 mM H₃PO₄ in acetonitrile; the temperature was 20 ° C; the sample volume was 100 ul; the gradient conditions were as follows: <img img-content="tb" img-format="tif" he="31" wi="166" file="00850001.tif" />the DAD settings were as follows:

<img img-content="tb" img-format="tif" he="20" wi="166" file="00850002.tif" />

EXAMPLE 18

HPLC assay for glutathione in PRBC and PRBC supernatant

0280the Sample was prepared as for the HPLC assay described above prepared. The HPLC conditions were as follows: analytical column = YMC ODS-AM-303, 250 mm x 4.6 mm, 5 .mu.m particles; guard column = Brownlee, 15 × 3.2 mm, 7 microns particles; the mobile phase for A was 10 mM H₃PO₄ in HPLC grade water; the mobile phase for C was 10 mM H₃PO₄ in acetonitrile; the temperature was 15 ° C; the sample volume was 75 .mu.l; the Gradient conditions were as follows: <img img-content="tb" img-format="tif" he="31" wi="167" file="00860001.tif" />the DAD settings were as follows:

<img img-content="tb" img-format="tif" he="15" wi="166" file="00860002.tif" />

EXAMPLE 19

A method for screening of adsorbents

0281A Test solution containing 25% plasma and 75% Erythrosol, was showing of the supernatant of PRBCs used. Erythrosol was as two separate parts of the initial solution (solution C and Solution D) many of which were sterilized separately. The final solutions were D prepared by mixing equal volumes of solution C and solution.

<img img-content="tb" img-format="tif" he="25" wi="123" file="00860003.tif" />

0282the Plasma Erythrosol solution was charged with 5 - [(β-carboxyethyl) amino] acridine added to a final concentration of 300 uM. Glutathione (reduced Form, Sigma Chemical Co.) was added to the mixture to a final concentration of 3 mM to obtain. Samples of adsorbents (0.2-0.8 g) were exactly in tarred 7 ml polypropylene vials with screw caps weighed (± 0.001 G). of the adsorbent, the required pre-wetting samples were suspended in 70% ethanol. The adsorbent was allowed to settle, and the supernatant has been removed. Residual ethanol by resuspending the zen adsorbent in distilled water to give Abset the adsorbent allowed, and skimming the supernatant away. The Adsorbensmassen were for the water content in the corrected calculation of the adsorption capacities. A 5.0 ml aliquot of the plasma / Erythrosol was added to each tube. The tubes were on a rotary device and taumelvermengt placed at room temperature for 24 hours. The vials were removed from the rotation device, and the adsorbent was allowed to settle. A sample of the supernatant was taken from each tube. The samples were centrifuged to remove the residual adsorbent. The samples were analyzed by HPLC to determine the residual amounts of 5 - [(β-carboxyethyl) amino] acridine analyzed. The above-described reverse phase assay was used to determine the residual amounts of glutathione used. The results of the adsorbent screenings are in Tables 7 and 8, respectively.

table 7 Adsorbent screening - Distance of 5 - [(β-carboxyethyl) amino] acridine from 25% plasma / 75% Erythrosol <img img-content="tb" img-format="tif" he="116" wi="166" file="00870001.tif" />

<img img-content="tb" img-format="tif" he="222" wi="166" file="00880001.tif" />

table 7 (continued) <img img-content="tb" img-format="tif" he="224" wi="166" file="00890001.tif" />

table 7 (continued) <img img-content="tb" img-format="tif" he="213" wi="166" file="00900001.tif" />

table 7 (continued) <img img-content="tb" img-format="tif" he="45" wi="166" file="00910001.tif" />

table 8th Adsorbent screening - Distance of glutathione in 25% plasma / 75% Erythrosol <img img-content="tb" img-format="tif" he="224" wi="167" file="00920001.tif" />

table 8 (continued) <img img-content="tb" img-format="tif" he="189" wi="167" file="00930001.tif" />

EXAMPLE 20

Adsorption of adsorbents

0283Adsorptionsisotherm experiments were used to determine the adsorption capacities (.mu.mol 5 - [(β-carboxyethyl) amino] acridine / g Adsorbent) for Various types of adsorbents made. In<figref idrefs="S134">17</figref> are for obtained different Ambersorbs adsorption isotherms compared for Adsorptionsisotherm for Purolite MN-200 demonstrated. were adsorption studies in 25% plasma / 75% Erythrosol solutions made, the 0.2-3 mM 5 - [(β-carboxyethyl) amino] acridine and from 0.6 to 10 mM GSH contained. The samples were incubated for 24 hours at room temperature shaken. The calculations under the application of the adsorption capacity of table 7 (22 mol / g) found that about 4 g would be required to Purolite MN-200 to acridine [(β-carboxyethyl) amino] - the amount of 5 in a 300 ml unit of PRBC from 300 uM to Endmenge to reduce by 1 uM. Less than 1 g of Ambersorb 572 (130 .mu.mol / g) would be required to a comparable to achieve distance. A similar Calculation came to the conclusion that less than 1 g of Ambersorb 572 would be required to the Amount of GSH in a 300 mL unit of PRBC from 6 mm to a Final amount of 500 uM in 150 ml of the supernatant reduce (50% HCT).

EXAMPLE 21

Flow studies Using multiple forms of activated carbon media

0284experiments were made to the removal of 5 - acridin [(β-carboxyethyl) amino] to investigate and GSH from PRBCs using a flow element. PRBCs (Erythrosol, glucose, 63% HCT) were at 300 uM a degradable 5 - [(β-carboxyethyl) amino] acridine derivative and 3 mM GSH metered, and at room temperature without shaking for 20 hours held before being exposed to the flow elements. the Flow media of the adsorption of the compound consisted of various forms of activated carbon, either as a composite Carbon / cellulose disk or as carbon fiber felt. the Media were included in a polycarbonate housing with 90 mm diameter (Cuno, Meridien, CT). The dose PRBCs were through the media at a flow rate of 5 ml / min pumped (Minipuls Gilson, Middleton, WI) and a PL 146 plastic container (Baxter Healthcare) collected. The results of this study are shown in Table 9 below.

table 9. Results of the flow study using different Activated carbon media <img img-content="tb" img-format="tif" he="101" wi="151" file="00950001.tif" />

EXAMPLE 22

Flow Verbindungsadsorptionselemente (CUNO media) gg. Batch Verbindungsadsorptionselemente (AQF media)

0285It Studies have been conducted, 5 - [(β-carboxyethyl) amino] acridine and GSH removal in flow gg. batch Verbindungsadsorptionselementen compared. The flow element consisted of a cellulose / Norit A Supra (Norit Americas, Inc. (Atlanta, GA)) - carbon disc, in a 90 mm housing was included. There were two different batch items used: one consisted of a fibrous Pica G277 activated carbon (AQF 500 g / m²); the other consisted of fibrous Purolite MN-200 (AQF 312 g / m²). Following the Dosage of 300 uM a degradable 5 - [(β-carboxyethyl) amino) acridine derivative and 3 mM GSH were the PRBCs by the cellulose / carbon medium pumped at a flow rate of 2 ml / min, after which the 5 - [(β-carboxyethyl) amino] acridine and GSH levels by 75 and 88% at 24 uM and 0.71 mM in PRBC supernatant fell off. The the flow element exposed PRBCs were then transferred to the 6 g-MN-200 charge element, which the 5 - [(β- carboxyethyl) amino] acridine and GSH levels reduced by another 5 and 1%, which after 24 Hours 20 uM and 0.67 mM reached. Exposing the PRBCs a 7 g Pica G277 batch element alone resulted to a decrease of 5 - [(β-carboxyethyl) amino] acridine and GSH levels by 92 and 54% at concentrations of 8 microns and 3 mm. Therefore, a passage was through the flow element in removing of GSH effective than the 24-hour Exposing the carbon batch element. The batch element alone however, was more effective at removal of 5 - [(β-carboxyethyl) amino] acridine than the combined flow and MN-200 elements. The flow by element seemed to be no deleterious effect on the PRBC ATP concentration to have. The K + quantities in the PRBCs goods by exposure to the flow element compared to a 24-hour exposure the carbon batch element low.

EXAMPLE 23

Long-term removal of degradation products

0286at this experiment, the 5 - [(β-carboxyethyl) amino] acridine and GSH levels in PRBCs investigated, wherein a fibrous PICA G-277 activated carbon element (AQF, 7.3 g, 500 m²/G) after 24 hours Exposure has been removed. This study was parallel to studies carried out in which the PRBCs exposed to continue an element were. <figref idrefs="S135">18</figref> shows that the concentrations acridine ((β-carboxyethyl) amino] - 5 of and GSH in the supernatant samples over in the absence of an element Storage durations of 1 to 4 weeks were significantly higher.

0287the acridine [(β-carboxyethyl) amino] - concentration of 5 was at 5 uM initially at ridden PRBCs after 35 days of Storage decreased in the presence of a removal member (MN-200). This shows that the removal of 5 - ((β-carboxyethyl) amino] acridine takes place in static storage conditions at 4 ° C.

EXAMPLE 24

Effect of the inclusion material (Membrane, woven, non-woven) to an IAD

0288the Use of an inclusion material surrounding the adsorbent medium, was for the primary Purpose of the particle retention examined. The primary Purpose is the particle retention. The membranes can mente but hemocompatibility of Ele increase, by the contact between the membranes and PRBCs is prevented. The membranes can readily modified with hydrophilic polymers (PEO, PEG, HPL) are to the hemocompatibility without change to increase the function. Approximately 10 g of the fibrous Pica G277 activated carbon medium (AQF 500 g / m²) were of a heat-sealed surrounded membrane, namely woven polyester or non-woven polyester material. PRBC units (300 ml) were treated with 300 uM a degradable 5 - [(β-carboxyethyl) amino] acridine derivative and 3 mM GSH dosed for 20 hours on a platelet shaker on held room temperature and then transferred to the IADs. The concentration of 5 - [(β-carboxyethyl) amino] acridine was monitored 24 hours. <figref idrefs="S136">19</figref> shows the 5 - [(β-carboxyethyl) amino] acridine levels in supernatant 300 mL PRBC units that had been exposed to the IADs consisting of fibrous Pica G277 activated carbon (500 g / m²) And included by a woven or non-woven membrane material. The PRBCs (Erythrosol, glucose, 62% HCT) were incubated with 300 uM of a degradable 5 - [(β-carboxyethyl) amino] acridine derivative and 3 mM GSH and metered before on a platelet shaker at room temperature the transfer shaken to the IADs. The PRBC-containing IADs were at room temperature for 24 hours shaken.

0289These Studies show that the woven Tetko-container the fastest removal kinetics for 5 - [(β-carboxyethyl) amino] acridine over 24 hours shows. The for all container materials after 2 weeks final levels achieved were similar, wherein the 5 - [(β-carboxyethyl) amino] acridine-concentrations to about 2 microns after 1 day declines, yet close to the day 8 again risen to 10 uM.

EXAMPLE 25

Effect of Verbindungsadsorptionselements the function of the red blood cells

0290indicators red blood cell function were about the course of the experiments for 5 - [(β-carboxyethyl) amino] acridine and GSH removal for different element configurations monitored. To the measured parameters included the percentage lysis, ATP and K⁺-Concentration. Table 10 shows that the ATP concentrations by the presence a connecting element removal generally not affected were: the decrease in ATP concentrations was approximately the same trol for Kon (No IAD) as for the MN-200 or Pica G277 elements. The amounts of K⁺ in PRBCs were found to ascending with time. The temperature, in which the distance was, influenced the K⁺-Quantities in the PRBC units, the connection distance elements over 20 days exposed away, not. Where the time duration of exposure was extended to 35 days, but varied the rate of increase K⁺ in PRBCs with the type of adsorbent, the final levels of 40 and 45 mmol / l for MN-200 or PICA G-277 elements were achieved, compared to the element-free control at 39 mmol / l. The percentage of red blood cells, the element in exposed and element-free control PRBC units was lysed was generally found with 0.1 and 1% after 24 hours. As shown in Table 11 and Table 12 and in <figref idrefs="S137">20</figref> and <figref idrefs="S138">21</figref> graphic shown, varying the% lysis with the type of adsorbent used significantly. Table 11 shows Lysewerte obtained as for PRBCs, the two Species were exposed immobilized adsorption elements for 35 days. Table 12 shows the Lysewerte obtained for PRBCs that in a woven polyester mesh for 42 days trapped loose adsorbent particles were exposed. A wrist-shaker was for the Dosing all PRBC units for used 1 minute, whereupon the PRBCs for 4 hours at room temperature were in a static state. The elements were for 24 hours on a platelet shaker at Room temperature held, whereupon it in for the duration of the study 4 ° C in a static state were. The immobilized MN-200 showed lower Hämolysemengen carbonaceous than immobilized PICA G-277, while the synthetic Ambersorb adsorbent one of the lowest when compared Hämolysemengen loose Partikeladsorbenzien revealed. The MN-200 showed a lower IAD hemolysis than the same non-immobilized adsorbent. Similar observations were for other IADs made in comparison with the same non-immobilized adsorbent.

table 10. ATP concentration (mol / dL) in PRBCs over time <img img-content="tb" img-format="tif" he="40" wi="167" file="00990001.tif" />

table 11. Percentage lysis at PRBCs (56% HCT), exposed to the various IADs, over time <img img-content="tb" img-format="tif" he="40" wi="166" file="00990002.tif" />

table 12. Percentage lysis at PRBCs (55% HCT), exposed to the various loose Partikeladsorbenzien, enclosed in a mesh <img img-content="tb" img-format="tif" he="59" wi="166" file="00990003.tif" />

0291the critical nature of the adsorbent with respect to the retention of red blood cell function illustrated. A comparative study of the effects of five different Adsorbents on the red Blutzellhämolyse shown. Ambersorb 572 yielded only 0.16% lysis at PRBCs (55%) after a 24-hour exposure, whereas Darco AC (Norit Americas, Inc. (Atlanta, GA)) 0.13% lysis revealed. These adsorbents were significantly better at minimizing hemolysis of red blood cells than the Purolite MN-200, Hemosorba CH-350 and Pica G277 adsorbents.

0292In Table 13 is shown a comparative study, in the supernatants of Sample of red blood cells, acridine [(β-carboxyethyl) amino] - glutathione and 5 contained, brought together with a number of different adsorbents were. Activated carbon adsorbents were the only type of adsorbent for sizable acridine [(β-carboxyethyl) amino] - reducing the concentrations of both 5 and glutathione was able. Both natural activated carbons (Norit and PICA) and synthetic activated carbons (Ambersorb) proved in the reduction of the compound to be effective.

table 13 properties of several different adsorbents <img img-content="tb" img-format="tif" he="108" wi="158" file="01010001.tif" /><!--footnotes removed--><!--footnotes removed--><!--footnotes removed--><!--footnotes removed-->

EXAMPLE 26

0293In This example illustrates the typical performance of an immobilized Adsorbenselements in a flow mode in plasma using described a Adsorbensmediums, composed of 30% Norit A Supra-carbon is (Norit Americas, Atlanta, GA) composed and produced by Cuno (Meriden, CT) as described previously in an earlier example.

0294the IADs were impregnated using a 30% Norit A Supra 90 mm Cuno R10SP medium in series with a Memtec-Hydroxypropylcellulosebeschichteten Polysulfone membrane 90 mm in diameter composed with 5 micron pores.

0295to completion the disposable product, a 1 l PL2410 bag on a 1/8 inch OD connected piping system and the piping system then the End of the IAD so connected that the distance from the center line the IAD to bag 40 cm. A cable system in the same size was attached to the inlet of the SRD so that when the irradiation bag was connected to it, the distance from the center line of the bag 30 cm.

02965 - [(β-carboxyethyl) amino] acridine was added to a final concentration of about 150 .mu.M 500 ml plasma, and the plasma was 6.4 J / cm² UVA for Inactivate pathogens irradiated. acridine [(β-carboxyethyl) amino] - The concentration of 5 after irradiation was about 82 uM.

0297table 14 shows measurements of clotting factor activity and 5 - [(β-carboxyethyl) amino] acridine-quantities after the treatment by the IAD, relative to values ​​before the flow by the IAD. The experiment was repeated three times. The mean values and standard deviations are given. Treatment duration was on average 14 minutes. It is apparent that almost no change the factors I, II, V, VII, X or measurements of the aggregate-clotting activity of prothrombin (PT) and activated partial thromboplastin time (aPTT) and very little change templates in factors XI and XII. Factors VIII and IX show something major changes, but these are acceptable, especially in the light of Regulation of recombinant proteins because of their lack of presence. The highly selective nature of the element should be noted, by almost back keeps the entire clotting activity, while 0.9% of 5 - [(β-carboxyethyl) amino] acridine the passage allowed.

table 14 <img img-content="tb" img-format="tif" he="58" wi="167" file="01030001.tif" />

EXAMPLE 27

0298The fibrous medium consisting of Dowex Optipore L-493 and attached to a non-woven polyester fiber matrix (Hoechst-L493) was prepared by the AQF-Division of Hoechst Celanese (Charlotte, NC). The performance of this Adsorbensmediums in a batch removal device was for platelets evaluated.

Platelet preparation

0299Single-donor apheresis platelet units the 3.5-4.5 × 10¹¹ platelets in 300 ml of 35% autologous plasma, 65% PAS III contained, were obtained from the Sacramento Blood Bank Center. 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen (Baxter Healthcare) each platelet unit was added to a Final concentration of 150 microns to reach. The platelet units (4-5) were pooled in a single PL-2410 plastic container and mixed thoroughly. The platelet pool was equally divided into 4-5 1L PL2410 plastic containers, each containing about 300 ml of the platelet mixture contained. The units were 3.0 J / cm² UVA photochemical treated and transferred to the corresponding removal member for the study. All experiments included a control platelet unit which treated photochemically (150 uM Psoralen, 3.0 J / cm² UVA), but not with a Entfernungselment been brought together.

Element preparation

0300Standard distance elements 2.5 g Dowex XUS-43493 contained, were by the Baxter Healthcare Corporation produced (Lot FX1032 D96F20042R). Experimental IADs were prepared with Hoechst-L493 media (Hoechst Celanese Corp.) which were delivered as a roll stock. The medium was measured and cut to the appropriate adsorbent for each IAD (5.0 g and 7.5 g) to obtain. The cut medium was placed in bags by pulse welding of 30 micron polyester mesh (Tetko) were constructed. The medium containing mesh bag were autoclaved (121 ° C, 20 min) and in sterile PL introduced 2410 plastic container. Alternatively, the mesh bags were in PL 2410 containers introduced, the container welded and the entire assembly by means of gamma-irradiation at 2,5 Mrad (SteriGenics, Corona, CA) sterilized. Excess air was from the elements using a syringe prior to transfer of the photochemically treated platelets manually evacuated.

adsorption kinetics

0301in the Following the photochemical treatment with 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen + UVA was the platelet mixture in PL2410 plastic containers transmitted with distance elements. The elements were on a standard Blutplättcheninkubator placed (Helmer) and at 70 Cycles / min shaken at 22 ° C. the platelet mixture samples were dissolved in 1-hour intervals for Located at the first 8 hours of storage. These samples were at 4 ° C stored and later to the rest of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen analyzed by HPLC analysis. The assay involved an initial Sample preparation which lyses the platelets and the 4 - 8-trimethylpsoralen '(4-amino-2-oxa) butyl-4,5', and free photoproducts unlocks. The supernatant of the sample preparation was on a C-18 reverse phase column with a gradient of increasing methanol KH₂PO₄Buffer analyzed.

In vitro platelet function

0302the platelet mixtures was shaken in contact with the removal of elements for 7 days. At a Study the platelet mixture with the IAD for contacted 24 hours, and under in sterile 1 L PL2410 plastic containers Using a sterile line welder (Terumo SCD 312) transferred. platelets were in the Blutplättcheninkubator brought back and for the remainder of the 7-day storage period stored.

0303After 5 or 7 days of storage was the platelet count and the pH for each platelet unit certainly. The pH and pO2 / pCO2 were measured using a Ciba-Corning Model 238 blood gas analyzer measured. The platelet count each Sample was determined using a Baker 9118+ hematology analyzer.

0304Several Assays were performed for the evaluation of in vitro platelet function. the shape change Platelet samples was monitored using a Chrono-Log model 500 VS Vollblutaggregometers. The change in shape was defined as the ratio the maximum change in the light transmission following the addition of ADP relative to the change in in the light transmittance in the Connection quantified to the addition of EDTA.

0305the Reaction of platelets to hypotonic stress was the Hypotonic Shock Response (HSR) assay evaluated. The change in the light transmission following the addition of a hypotonic solution was prepared using a Chrono-Log model 500 VS Vollblutaggregometers measured. the Data are presented as percent recovery from hypotonic stress two minutes after the extinction had reached its minimum value, reported.

0306the aggregability platelet in response to ADP / Collagenagonist was the change in optical transmittance specify how using a Chrono-Log model 500 VS Vollblutaggregometers measured.

0307Of the Status of platelets was evaluated by scoring the platelets. The samples were blinded and morphology-point values ​​for 100 platelets for each sample added. The highest possible score is 400 (washer = 4, intermediate = 3, ball = 2, Dendrite = 1, balloon = 0).

0308the Platelet activation as indicated by expression of p-selectin (Granular Membrane Protein, GMP-140) was measured. CD62 antibody was for the test sample used mouse control IgG1 was used for the negative control, and CD62 antibodies with Phorbalmyristatacetat (PMA) was used for the positive control. The samples were analyzed on a FACScan (Becton Dickinson). The percentage activation which is reported is, relative to the positive control indicated and represents the difference between the test value and the negative control value represents.

adsorption kinetics

0309the Effect of Low Adsorbenskügelchen in a fiber matrix was investigated. Platelet units the 4.0 × 10¹¹ platelets in 300 ml of 35% plasma, 65% PAS III containing, were treated with 150 uM 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen + 3.0 J / cm² UVA treated. The IADs were Hoechst-L493 with an adsorbent loading of 450 g / m² prepared. The IADs were sterilized by gamma irradiation. In connection the treatment with psoralen + UVA, the platelets were added to transfer the distance elements. The samples were dissolved in 1-hour intervals and extracted for residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen analyzed by HPLC. In<figref idrefs="S139">22</figref> will the kinetics of removal of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen for IADs compared that contained 5.0 g and 7.5 g Hoechst-L493 media, to those of a standard distance element, 2.5 g of loose beads contained. The IADs contained either 5.0 g Hoechst-L493 (triangles) 7.5 g Hoechst-L493 media (squares) or 2.5 g loose Dowex L493-Adsorbenskügelchen (Circles). The Hoechst-L493 media containedAdsorbenskügelchen at a loading of 450 g / m²,

0310the Kinetics of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethyl psoralen adsorption were for Hoechst media when compared to an equivalent mass of loose Adsorbenskügelchen slower. The distance element 2.5g loose beads containing achieved the lowest levels of residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen in short periods (1 hour). For longer periods but showed the IADs containing 7.5 g and 5.0 Hoechst-L493 media, better performance. In this study, the adsorption kinetics were in all three segments Entfernungsele relatively quickly by the Target of &lt;0.5 uM of residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen achieved in less than 4 hours of contact.

0311one Note that longer at all Durations (6-8 Hours) the Hoechst-L493 IADs containing a greater mass of adsorbent, lower amounts of residual 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen achieved. This observation suggests that the slower adsorption kinetics for the Hoechst-L493 IADs a series of restrictions mass transport (liquid stream . Diffusion) is payable and not the result of loss of functional adsorbing due to fiber bonding. Other studies show that Hoechst-L493 media with lesser amounts of adsorbent loading (200-300 g / m²) of the liquid an easier penetration of the media allowed, leading to faster Adsorption kinetics lead. previous studies (Data not shown) with Hoechst medium containing Amberlite XAD-16 adsorbent at a loading of 160 g / m² contained, revealed that the adsorption kinetics by incorporating into a fiber matrix were not affected at a low loading amount.

Platelet yield and in vitro platelet function

0312In This section presents two separate studies the data. The platelet units within each study were from a single pool, so that the Effect of the IAD media format, the adsorbent and the contact time could be clearly evaluated.

0313In The first study was the effect of the fiber formation (Hoechst media) on the yield and platelet function following a extended contact evaluated (5 and 7 days). Total 4 platelet units derived from a single pool, were used in this study. The IAD-free control unit has been treated with psoralen + UVA. Two of platelet units were combined with 5.0 g Hoechst-L493. One unit was with the IAD for brought together 24 hours before they transferred to an empty PL 2410 storage container has been. The other unit remained in contact with the IAD for the duration the study. The Hoechst-L493 IADs were sterilized with steam (120 ° C, 20 min). The standard distance element (2.5 loose Dowex XUS-43493), which was obtained from Baxter, was sterilized by gamma irradiation. It is noted that the platelets do not the standard distance element following an 8- to 16-hour contact have been removed, as for execution used with the element typically loosely adsorbent.

0314the Results of platelet count and in vitro function following 5 and 7 days of storage are given in Table 15A and Table 15B together.

table 15A. (Day 5) Comparison of Hoechst-L493-fibrous media with standard distance element platelet yield and in vitro function following 5 days storage <img img-content="tb" img-format="tif" he="116" wi="167" file="01080001.tif" />

table 15B. (Day 7) Comparison of the Hoechst-L493-fibrous media with standard distance element platelet yield and in vitro functional after 7 days storage <img img-content="tb" img-format="tif" he="116" wi="167" file="01090001.tif" />

0315the platelet yields for the Hoechst-L493 IADs (5.0 g) were considerably better than the yield for the default removing member (2.5 g). Loss of &lt;10% were at 7 days Storage in continuous contact with the Hoechst-L493 IAD (5.0 g) achieved. Both platelet units treated with the Hoechst-L493 IADs showed better Power in the shape change, Aggregation and GMP-140 assays as the IAD-free control. the platelets, in contact with the Hoechst-L493 IAD (5.0 g) for the entire remained five days, showed comparable in vitro function the platelets, after 24 hours Beam Contact were. Interestingly, the platelets in contact with the Hoechst-L493 IAD remained, better performance in GMP-140 assay. the Difference in power between the two was even after 7 days even bigger. These Obs respectfully suggests that the contact with the IAD rate of the p-selectin expression diminished by the platelets during storage.

0316the second study looked IADs containing 5.0 g and 7.5 g Hoechst-L493 media contained, in order to determine whether a significant decrease in platelet yield or carried out in vitro function with a higher mass of medium. In this study, the Hoechst-L493 IADs were by gamma irradiation sterilized. A standard distance element (2.5 g Dowex XUS-43493) was included in the study. remained Platelets in contact with the removal of elements for the entire duration of the study. The results of the platelet count and the in vitro functional after 5 and 7 days Storage are summarized in Table 16A and Table 16B.

table 16A. (Day 5) Evaluation of gamma sterilierten Hoechst-L493-fibrous media platelet yield and in vitro function following 5 days Storage (no transfer) <img img-content="tb" img-format="tif" he="103" wi="166" file="01100001.tif" />

table 16B. (Day 7) Evaluation of gamma sterilierten Hoechst-L493-fibrous media platelet yield and in vitro functional after 7 days Storage (no transfer) <img img-content="tb" img-format="tif" he="104" wi="166" file="01110001.tif" />

0317the Results, which are summarized in Tables 16A and 16B are, the results of similar, the were obtained in the first study. The platelets with the Hoechst-L493 IAD were contacted gave significantly higher yields than the platelets, with the standard distance element were contacted. The platelet yield was slightly lower for the 7.5 g Hoechst-L493 IAD relative to 5.0 g-IAD. Platelets, which had been treated with the Hoechst-L493 IADs showed comparable performance to the control platelets in all in vitro assays. Platelets, which had been contacted with the Hoechst-L493 IADs showed improved performance in the aggregation assay relative to the IAD-free Control on day 7. The GMP-140 assay has not been made on the day. 7

0318the prepared IADs that with Hoechst-L493 media (5.0, 7.0 g) had been, were superior in vitro function compared to standard elements Entfernungsele (2.5 g loose XUS-43493), which is in contact with the platelets for 5 days had been stored. About that also showed the platelets, with 150 uM of 4 '- (4-amino-2-oxa) butyl-4,5', 8-trimethylpsoralen + 3.0 J / cm² UVA treated, improved in vitro function as by shape change, Aggregation and GMP-140 assays displayed when using Hoecht-L493 IADs (5.0 g) for contacted 5 and 7 days. An additional study in which a 5- to 7-day Storage of platelets (No psoralen / UVA) was compared with and without IAD (5.0 g Hoechst-L493) demonstrated that storage with an IAD platelet power can be improved, as indicated by in vitro functional assays.