Single-use manifold for automated, aseptic transfer of solutions in bioprocessing applications

ES 2 319 635 T3

DESCRIPTION

Single-use manifold for automated aseptic transfer of solutions in bioprocess applications.

Field of the invention

The invention relates generally to the aseptic transfer of solutions from one or more storage or dispensing containers for fluids and / or biological processes. Disposable collecting systems perform the transfers required in bioprocess applications. Automated dispensing is achieved with the invention, preferably in association with one or more remotely controlled pinch valves.

Background of the invention

Good manufacturing practices and government regulations are at the core of any pharmaceutical, biotech and biomedical manufacturing process or procedure. These manufacturing processes and procedures, as well as the associated instrumentation, must pass mandatory, often time-consuming and costly validation processes.

For example, the instruments used for the separation and purification of biomedical products, for obvious reasons, must meet strict cleaning requirements. Cleanliness validation of old or new purification instruments (such as instruments for preparative chromatography or tangential flow filtration) may require up to 50 scratch tests of exposed surfaces and subsequent bioassays of scratches. For each piece of purification instrumentation, the associated and recurring cost of each cleaning validation can easily exceed several thousand dollars.

To reduce such costs and cleaning validation expenses, and / or to reduce the times when cleaning is needed or mandatory, the pharmaceutical and biotechnology industries increasingly use collapsible and disposable plastic tubes and plastic bags, pre-sterilized for transfer and storage of solutions. Sterilization is accomplished by subjecting the entire assembly of tubes and bags to gamma irradiation, or to an atmosphere of ethylene oxide. Assembled sets of tubes and bags thus pre-sterilized, aseptically packaged, are commercially available (currently from TC Tech; HyClone; St Gobain Performance Plastics, for example) and are used for manual transfer of solutions. Typically, the solution transfer procedure requires a technician to operate a peristaltic pump and manually open and close clamps on the tubes to direct the solutions from the reservoirs to the storage bags. While this procedure reduces cleaning efforts and cleaning validation expenses, it still requires operator time and interaction, and the consistent accuracy and precision of these methods is dependent on operator experience.

Dispensing procedures characterized by automation (which may include sensors, monitors, and programmable controllers) are common knowledge. Keys et al., In US Patent No. 5,480,063 and US Patent No. 5,680,960, describe fluid dispensing units that control fluid volumes in combination with a closed loop system, which, according to those patents, allows to avoid the use of vents. The fluid to be dispensed exits the closed-loop apparatus through a fill tube, operated by a controller. These methods do not take into account the cleaning needs or the costs and expenses of cleaning validation, necessary if this type of systems are to be used in the pharmaceutical and biotechnological industries for the dispensing, conduction, combination or separation of biological or chemical fluids. European patent EP 1236644 (prior art in the sense of Article 54 (3) and (4) EPC) describes an apparatus for dispensing liquid, eg for pharmaceutical applications, from one container to another. The apparatus has a syringe connected to a distributor, which in turn is connected by means of tubes to various containers. The syringe is adapted to draw liquid from a first container and pump it into a second container. The connections between the syringe and the various containers are opened and closed by means of pinch valves located in the tubes that connect the dispenser to the containers. There may be a filter between a destination container and the outlet of the tube that connects the container to the distributor.

Previous systems may incorporate diaphragm valves, in direct contact with the solution in process, and such valves are a potential source of contamination. Thus, diaphragm valves require expensive cleaning validation procedures.

It has been found that, by proceeding in accordance with the present invention, significant cost savings and better efficiency can be obtained in a system incorporating aseptic automated collectors within the technological field comprising plastic tubes and pre-sterilized plastic bags for use. unique, having at least one folding part. Each of the components that contact the biological or chemical fluid are pre-sterilized and disposable after use.

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Summary of the invention

The present invention is directed to collecting units that are pre-sterilized and disposable, becoming sterilized and packaged single-use units to be used directly from the warehouse, and thus eliminating long and tedious cleanings and tests in the place where they are to be used. be used. Various embodiments are described. They all consist of lengths of tubing and various disposable storage or collection bags, each of which has various inlet and / or outlet conduits that can be selectively opened and closed. The tube sections interact with one or more pinch valves that are remotely operated. Remote management is automated by means of a programmed controller to carry out the procedures according to the chosen embodiment.

The general objective of the present invention is to provide improved disposable collection systems for the automated aseptic transfer of solutions in bioprocess or chemical process applications.

Another object of the present invention is to provide improved apparatus combining the use of pinch valves with sterile, disposable dispensing manifold units.

Another object of the present invention is to provide improved apparatus that greatly reduce the consumption of time and resources, intended for cleaning procedures for transfer equipment used in the pharmaceutical and biological industries and in laboratories in which the contamination cannot be tolerated. contamination of biological and / or chemical fluids.

One aspect of the present invention is to reduce the need for validation procedures for equipment used in the separation and purification of fluids such as those for the preparation, separation and dispensing of biomedical products.

Another aspect of the present invention is that it meets the cleaning needs for procedures such as fluid dispensing, preparative chromatography and tangential flow filtration, while automating the corresponding operations.

These and other objectives, aspects, features, improvements and advantages of the present invention will be clearly understood by taking the following detailed description into consideration.

Brief description of the figures

Throughout this description, reference will be made to the attached figures, in which:

Fig. 1 is a somewhat schematic illustration of a presterilized disposable system that is especially suitable for transferring and collecting solutions;

Fig. 2 is an illustration of the disposable system of Fig. 1 in operative association with pinch valves, at least one of which is remotely controllable;

Fig. 3 is an illustration of the combination of the features of Fig. 1 and Fig. 2, shown together with means for use in transferring solutions through the system;

Fig. 4 is a somewhat schematic illustration of a presterilized disposable system that is especially suitable for use in automated preparative chromatography;

Fig. 5 is an illustration of the disposable system of Fig. 4 in operative association with pinch valves, at least one of which is remotely controllable;

Fig. 6 is an illustration of the combination of the features of Fig. 4 and Fig. 5, shown together with means for use in transferring solutions through the system;

Fig. 7 is a somewhat schematic illustration of a presterilized disposable system that is especially suitable for automated tangential flow filtration procedures;

Fig. 8 is an illustration of the disposable system of Fig. 7 in operational association with pinch valves, at least one of which is remotely controllable; Y

Fig. 9 is an illustration of the combination of the features of Fig. 7 and Fig. 8, shown together with means for use in transferring solutions through the system.

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Description of the individual realizations

In Figs. 1-3 illustrates a system specially designed for use as a system for the automated and aseptic transfer of solutions. The fluids processed in accordance with this invention are referred to interchangeably hereinafter as biotech fluids, pharmaceutical fluids, chemical fluids, etc. They are understood to be solutions, liquids, systems containing gases, and the like. In general, they are hereinafter referred to as biotech fluid or fluids.

In the pharmaceutical and biotechnology industries, media preparation departments typically prepare the solutions used in a solution production protocol that follows good manufacturing practices. Media preparation departments are responsible for maintaining dissolution recipes, preparing and storing buffers, and other tasks that require consistency and precision. For example, buffer solutions are prepared in large vats, and then pumped through sterilizing filters, for example one having a porosity of 0.1 µ. Typically, these solutions are filled with disposable presterilized bags for later use. A media preparation department may also be responsible for providing inoculation solutions to bioreactor operators. At the conclusion of a bioreactor batch, the reactor broth is often stored in sterile storage bags for further processing.

Fig. 1 shows the pre-sterilized disposable components of the invention. In general, these disposable components are a manifold assembly with transfer tubes and various bags. Various disposable storage / collection bags are shown, 21,22,23. Each has three connecting tubes. The primary inlet tube consists of an aseptic connector 24 and a manual closing clamp 25, all of generally known construction. During storage of the solution, the aseptic connector is covered with an end cap (not shown) to protect the connector 24 from contamination. The manual closing clamp 25 is closed during storage of the solution. These components are shown in a first connecting tube 30.

The second connection tube 26 consists of a short length of tube connected to the bag, with a manual closing clamp in the closed position. This tube and clamp assembly is used to relieve gases and / or pressures generated inside the bag during the filling operation. The third connection tube 27 is identical to the second connection and consists of a short length of tube and a clamp. Thus it can be used as an auxiliary inlet and / or outlet for the recirculation of the bag's content.

During a typical bag filling operation, the first and / or last collection bag can serve as a quality control bag. Typically these QC bags will be smaller in volume, eg one liter. During the initial priming cycle of the system, the first of these quality control (QA) bags is filled with the in-process solution. At the end of the refill cycle, when all the bags, normally larger in volume than the QA bags, have been filled with the product of the operation, the second QA bag is refilled. The solutions contained in the QA bags are then analyzed for possible contamination or other required QA data.

After completing the bag filling process, the manual closing clamps on each bag are closed and the aseptic connecting tubes are disconnected. During storage the connectors are protected by terminal caps (not shown).

Referring to the sterile disposable assembly of a manifold with transfer tubes of Fig. 1, one of these units is generally shown at 28. This represents a generalized manifold for automated transfer of solutions. The inlet end section 29 of the transfer tube 31 of the unit 28 is for communication with a solution container, eg a tank, typically of sterile solution. The sterile assembly of a manifold with transfer tubes 28 is shown with an optional in-line pressure sensor 32 and a disposable sterilization filter 33. The filter outlet tube 33 has the end tubes connected in series. By means of a suitable device that is in charge of transferring it, the solution travels from the tank or tank through the sensor 32 (if any) and the filter 33, and then it is directed through the final tubes in series to the sterilized disposable bags of storage.

Fig. 2 shows a set of pinch valves 41, 42, 43 and their respective relative positions with respect to the storage bags. Some or all of these valves can be remotely controlled, and are typically electrically or pneumatically actuated. A typical assembly accommodates up to twelve or more pneumatically controlled pinch valves. The same number of storage bags can be accommodated. Fig. 2 shows the relative positions of the clamp valves in association with the optional pressure sensor and disposable sterilization filter. Fig. 3 shows the relative position of the manifold and transfer tube assembly 28 with the tub 44 and piston of a pump unit 45. Preferably, the pump is a low-friction, high-precision peristaltic pump that provides full filling. fast and reproducible bags. An example is the Watson Marlow 620 RE peristaltic pump.

Access to the storage bags is facilitated through the pinch valves. Pinch valves are normally closed and typical pneumatic pinch valves require pressurized air (for example 80-100 psi) to open. When this pinch valve is pressurized, the solution is allowed to enter the storage bag.

ES 2 319 635 T3 while the air in the bag escapes through the filter of the integrated bleed valve. Pinch Valves are pneumatically or electrically controlled pinch valves (currently supplied by ACRO Associates, Inc.). They are installed on the outside of the tubes and are controlled by a multi-valve controller (currently supplied by SciLog Inc.) or other computer-based process control logic device (PLC). External clamp valves guide the solution into the manifold without affecting the sterile environment inside the tubes. The diaphragm valves used in other systems are in constant contact with the solution in process, whereas the pinch valves do not have contact with the solution in process.

The optional disposable pressure sensor 32 constantly measures the back pressure of the filter. This sensor can supply information to an appropriate controller to avoid undesirable events. For example, a controller may sound an alarm when a user-set safety pressure limit is exceeded, indicating that the capacity of the sterilant filter has been exhausted. Details on this aspect are found in US Patents 5,947,689 and US 6,350,382, and in US Patent Application Publication 2002/0043487.

The controller can be a standalone unit or be associated with another device. In a preferred arrangement, the controller is associated with the pump unit 45. This is shown as 46 in Fig. 3. The controller controls the operation of the remote control valves, whatever their shape. The speed of filling the batches as well as the volume of the batches delivered to each storage bag can be programmed by the user through the control program integrated in the controller. The controller provides automatic filling of bags by volume, by weight, or based on fill time and pump flow.

Typically, a user-created program will be implemented for the automatic filling of the storage bags according to Figs. 1-3. This is described for a SciPro controller from Scilog, Inc., generally described in US Patents 5,947,689 and US 6,350,382, and in US Patent Application publication 2002/0043487. These methods prevent the generation of excess pressure, as well as associated leaks and bag failures. For example, if so programmed, the controller will completely stop pumping when the safety pressure limit set by the user is exceeded.

An example of a solution transfer program to control the manifold is as follows. In SciPro edit mode, the user types and saves a multi-bag measurement program. The following is an example of a simple program to fill three 20 liter bags, bags 21, 22 and 23.

Example of filling program

<td> 000</td><td>START</td><td>The following steps of the program are typed in edit mode</td>

<td></td><td></td><td></td>

<td> 001</td><td>CW</td><td>The motor runs in the direction schedule</td>

<td> 002</td><td>RUN</td><td>The engine starts</td>

<td> 003</td><td>V 100000</td><td>Pinch valve 41 opens, other valves close</td>

<td> 004</td><td>RATE: 5.0 1 / min</td><td>Pump flow 5 liters per minute</td>

<td> 005</td><td>TIME: 00:04:00</td><td>The pump runs for 4 minutes, the Bag 21 is filled with 20 Liters</td>

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<td> 006</td><td>STOP</td><td>The pump stops</td>

<td> 007</td><td>V 020000</td><td>Pinch valve 42 opens, other valves close</td>

<td> 008</td><td>TIME: 00:00:02</td><td>2 seconds of standby time</td>

<td></td><td></td><td></td>

<td> 009</td><td>RUN</td><td>The pump starts</td>

<td> 010</td><td>RATE: 5.0 1 / min</td><td>Pump flow 5 liters per minute</td>

<td> 011</td><td>TIME: 00:04:00</td><td>The pump runs for 4 minutes, the Bag 22 is filled with 20 Liters</td>

<td></td><td></td><td></td>

<td> 012</td><td>STOP</td><td>The pump stops</td>

<td> 013</td><td>V 003000</td><td>Pinch valve 43 opens, other valves close</td>

<td> 014</td><td>TIME: 00:00:02</td><td>2 seconds of standby time</td>

<td></td><td></td><td></td>

<td> 015</td><td>RUN</td><td>The pump starts</td>

<td> 016</td><td>RATE: 5.0 1 / min</td><td>Pump flow 5 liters per minute</td>

<td> 017</td><td>TIME: 00:04:00</td><td>The pump runs for 4 minutes, the Bag 23 is filled with 20 liters</td>

<td> 018</td><td>STOP</td><td>The pump stops</td>

<td> 019</td><td>V 000000</td><td>All pinch valves close</td>

<td></td><td></td><td></td>

<td> 020</td><td>COUNT: 1</td><td>Program steps 000 to 020 are executed only once</td>

<td> 021</td><td>EMD l</td><td> _____________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________1</td>

By changing the RATE (flow) and TIME (time) program steps, the program is adjusted to any volume of the storage bags. Additional “RUN” blocks (pump running) can be inserted in the program to increase the number of bags to be filled (up to 12, in the example). However, an analogous software program can be generated in which the storage bags are filled based on the VOLUME (volume) or WEIGHT (weight) program commands. To fill the bags based on weight, a scale of appropriate capacity is required. An optional scale or load unit 47 can be provided to supply data to the controller in this case. It is noted that this embodiment measures user defined fluid volumes, then automatically switches to the next empty storage bag to be refilled.

A second embodiment, which is generally illustrated in Figs. 4-6, perform automated preparative chromatography. In preparative chromatography, the in-process solution containing the biomolecule of interest is pumped through a column of gel-like particles (stationary phase) suspended in a liquid. The biomolecule of interest specifically interacts (eg, through ion-ion interactions, hydrophobic interactions, size exclusion, or affinity) with the stationary phase, thus slowing the progression of the biomolecule through the column. Ideally, other dissolved biomaterials will only weakly interact with the stationary phase, thus rapidly exiting the column.

The result is the concentration, as well as the separation, of the biomolecule from the rest of the matrix of the solution in process. The introduction of an elution buffer will change the local chemical environment of the stationary phase, causing the release of the biomolecule, which can thus be collected outside the column in a relatively small volume of elution buffer.

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In automated preparative chromatography, the column containing the stationary phase is first washed and / or equilibrated with an appropriate buffer solution. This wash and / or equilibration cycle is followed by a loading cycle during which the in-process solution is pumped through the column. The biomolecule of interest adheres to the stationary phase. The loading cycle can take many hours, depending on the volume of the solution in process and the pumping rate at which the solution is pumped through the column. The loading cycle is followed by a second wash cycle to remove any non-adsorbed biomaterial from the column.

An elution buffer is then introduced to remove the biomolecule from the column. This removal of the biomolecule is carried out by means of a step gradient or a linear gradient. After peak collection is complete, the chromatography column is regenerated and rebalanced using appropriate buffers as is generally known in the art.

The assembly of the manifold and transfer tubes 48 represents a generalized manifold for automating preparative chromatography procedures. In operation, and using the controller system, the pinch valve 51, pneumatically controlled in the example in the example, is pressurized and therefore opens, thereby allowing access from the bag 54 of the washing and / or balancing buffer. With a user-definable pump rate, the wash buffer is pumped through an in-line disposable pressure sensor 55, through a bubbler (not shown), through chromatographic column 56, and through from an ultraviolet (UV) flow cell or detector 57. At the outlet of the flow cell, the wash / equilibration buffer is collected in a waste container or bag 58 while the clamp valve 49 is pressurized and therefore open .

During the load cycle, the pinch valves 51 and 49 are open / pressurized, while the pinch valves 52, 53 and 59 remain closed. Pump unit 45 pumps the in-process solution through collecting system 48, column 56, and flow cell 57 and is collected in waste container or bag 58. In some chromatography applications, the in-process solution emerging from flow cell 57 needs to be separately stored in a "processing receiving bag" (not shown) for possible reuse. Another pinch valve (not shown) would give access to said "processing receiving bag".

The loading cycle is followed by a flush cycle (valves 51 and 49 open / pressurize, the rest of the pinch valves are closed) which carries any non-adsorbed material from the column to the waste. By opening pinch valves 53 and 49, the elution buffer from bag 63 is introduced into the column and initially pumped to the waste. However, when the signal from the UV detector 57 exceeds the user-defined value, the clamp valve 59 opens, thereby giving access to the spout collection bag 61, while the valve 49 closes. At the end of peak elution, valve 59 closes again and at the same time valve 49 opens.

After the material of interest has been collected in bag 61, the chromatographic column 56 must be regenerated and rebalanced. The column regeneration process is easily automated through access to appropriate buffers (not shown), which are generally known in the art. Depending on the underlying chromatographic complexity of the application, access to five or six buffers may be required, which can be supplied in their own disposable bags, if desired. Similarly, if multiple peaks of product are to be collected, additional peak collection bags, as well as additional pinch valves, may need to be incorporated into the manifold assembly and transfer tubes 48.

The pre-sterilized disposable components of the manifold and transfer tube assembly 48, shown as a feed section, and a second bag and tube assembly 64 for the chromatographed fluid are shown in Fig. 4. Each of the individual storage bags / Pickup 54, 62, 63 shown in Fig. 4 has three connecting tubes. The primary inlet tube 65 consists of an aseptic connector 66 and a manual closure clamp 67. During storage of the solution, the aseptic connector is covered with an end cap to protect the connector from contamination. The manual locking clamp is closed during solution storage.

The second bag and tube assembly 64 consists of a short length of tube 68 connected to the bag with a manual closure clamp 69 in the closed position. The second tube and clamp assembly 71 is used to relieve gases and / or pressures generated within the bag during the filling operation. The third connecting tube 72 is identical to the second tube and clamp assembly 71 and is used as an auxiliary inlet and / or outlet for recirculation of the contents of the bag.

The disposable storage / collection bags 58 and 61 are connected to the remainder of the tubing assembly 72 as shown in Fig. 4 and Fig. 5. Fig. 5 shows the relative positions of the pinch valves 51, 52 , 53, 49 and 59 and the position of the pressure sensor 55. Fig. 6 shows the insertion of the manifold tube to the peristaltic pump 45 as well as the connections to the chromatographic column 56 and the detector 57.

In a typical chromatographic application, disposable storage bags 54 (for wash buffer) 52 (for in-process dissolution) and 63 (for elution buffer) have been prefilled, for example using the embodiment of Figs. 1-3. When the chromatography is complete, the manual closing clamps of each of the collection bags are closed, 58 (for waste), 61 (for the collected peak) and that of the solution in process (if desired, not shown), and the aseptic connecting tubes are disconnected. During storage, the ends of the aseptic connecting tubes are protected with end caps.

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In additional reference to the SciPro controller programmed to control the manifold mounting for chromatography, the corresponding mode allows a sequence of simple commands to be typed and stored, e.g. For example, RUN (running), RATE (flow), TIME (time), VOLUME (volume), P LIMIT 1 (positive slope of signal 1) and valve statuses such as V = 000000 (all valves of clamp are closed) or V = 123456 (all clamp valves are open).

This controller mode is organized into applet blocks. The termination declaration of a program block can be a declaration of the type “vOlUME”, “TIME”, “P LIMIT D1 (or D2)” or “N LIMIT D1 (or D2)”. The statement "P LIMIT D1 = 5%" means "positive slope of the D1 detector signal with a threshold value of 5% of full scale (FS)". See Example Chromatography Program.

Chromatography Program Example

<td> 000</td><td>START</td><td>Start of 1^er wash cycle</td>

<td> 001</td><td>CW</td><td>Direction of motor clockwise</td>

<td> 002</td><td>RUN</td><td>The engine starts</td>

<td> 003</td><td>RATE 0.25 L / M</td><td>Pumping flow during wash cycle</td>

<td> 004</td><td>V 100050</td><td>Wash buffer 51 led to Waste 49</td>

<td> 005</td><td>VOLUME 1.0 Liters</td><td>4 Minutes, end of 1^er wash cycle, total volume TV = 1.0 L</td>

<td></td><td></td><td></td>

<td> 006</td><td>RATE 1.00 L / M</td><td>Load flow, start of charge cycle</td>

<td> 007</td><td>V 020050</td><td>Dissolution in process (52) led to Waste 49</td>

<td> 008</td><td>TIME: 00:02:00</td><td>2 Minutes, end of charge cycle, TV = 3.0 L</td>

<td></td><td></td><td></td>

<td> 009</td><td>RATE 0.25 L / M</td><td>Start of 2^or wash cycle</td>

<td> 010</td><td>V 100050</td><td>Wash buffer (51) led to Residues (49)</td>

<td> 011</td><td>VOLUME 1.0 Liter</td><td>4 Minutes, end of 2^or wash cycle, TV = 1.0 L</td>

<td></td><td></td><td></td>

<td> 012</td><td>V 003050</td><td>Elution buffer (53) led to Residues (49)</td>

<td> 013</td><td>P LIMIT DI = 5%</td><td>Threshold value to detect the beginning of the collection of the peak volume</td>

<td></td><td></td><td></td>

<td> 014</td><td>V 003400</td><td>Elution buffer (53) led to Collection (59)</td>

<td> 015</td><td>N LIMIT DI = 10%</td><td>Threshold value Di, end of picking up the peak volume</td>

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<td></td><td></td><td></td>

<td> 016</td><td>V 003050</td><td>Elution buffer (53) led to Residues (49)</td>

<td> 017</td><td>VOLUME 1.0 Liter</td><td>Elution volume, end of elution, TV = 5.0 L</td>

<td></td><td></td><td></td>

<td> 018</td><td>RATE 0.50 L / M</td><td>Start of 3^er wash cycle</td>

<td> 019</td><td>V 100050</td><td>Wash buffer (51) led to Residues (49)</td>

<td> 020</td><td>TIME: 00:02:00</td><td>2 minutes, end of 3^er wash cycle, TV = 5.0 L</td>

<td></td><td></td><td></td>

<td> 021</td><td>STOP</td><td>The pump stops</td>

<td> 022</td><td>V 000000</td><td>All V-valves closed</td>

<td> 023</td><td>END</td><td>End of the program</td>

For example, on line 014, the SciPro changes from "Residual" to "Pickup" when the D1 signal has a positive slope and a value greater than 5% of full scale (line 013). The statement "N LIMIT D1 = 10%" means: "Negative slope of detector D1 signal with a threshold value of 10% of full scale". In line 016, the controller changes from "Pickup" to "Residual" when the signal from D1 has a negative slope (fall of the peak) and a value of 10% of the full scale (line 15).

The user can edit and / or modify the values ​​of: RUN (running), RATE (flow rate), TIME (time), VOLUME (volume), P LIMIT 1 (see above), N LIMIT D1 (see above) and the status of the valves at any time during the chromatographic operation. User-designed application programs can be uploaded or downloaded from an external computer at any time by using the computer's hypertherminal.

It can be appreciated that, with this embodiment, sequential scheduling of events is achieved. These include sequential programming of wash, load and elution cycles. The controller can initiate buffer selection, loading, and peak volume collection. Typical online concentration detectors can be Wedgewood UV and / or pH detectors, which have 4-20 mA output signals that can be monitored simultaneously. A typical pump is a Watson Marlow 620 R peristaltic pump capable of generating 60 psi (4.0 atm) at a pumping rate of 15 liters per minute.

User-defined detection threshold levels are used to switch valves and for peak volume collection. All solution handling parameters such as pump flow rates, column pressure and valve positions can be monitored and documented in real time and can be printed or archived electronically.

In a third embodiment, automated tangential flow filtration is carried out using a modified system designed for this use. The previously mentioned patents US 5,947,689 and US 6,350,382, and the publication of the US patent application 2002/0043487 disclose the automation processes of the tangential flow filtration (TFF). These are combined with the use of single-use disposable manifolds, which also include disposable pressure sensors and collapsible disposable storage bags, and the use of remotely controlled pinch valves.

A typical TFF application employing a presterilized disposable manifold is shown in Figs. 7-9. Fig. 7 shows the disposable presterilized components, including a filtered fluid tube section having a filtrate collection bag 81 as well as a working solution bag 82 within the flow filtration section of the tube. These components are aseptically sealed and in pre-sterilized (eg irradiated) packaging. At the start of the TFF application, the filtrate collection bag 81 is empty and deflated, and has been aseptically connected to the TFF manifold. The in-process solution bag was pre-filled, for example, using the system of Figs. 1-3. The process solution bag 82 is placed on an optional scale 83 and aseptically connected to the rest of the system. In some applications, information about weight may be supplied to the controller when running the control program.

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The presterilized components of this embodiment are shown in Fig. 7. The filtrate collection bag 81 has three connecting tubes. The primary inlet tube 84 consists of an aseptic connector 85 and a manual closure clamp 86. During solution storage, the aseptic connector is covered with an end cap to protect the connector from contamination. The manual locking clamp is closed during solution storage.

The second connecting tube consists of a short length of tube 87 connected to the bag with a manual closing clamp 88 in the closed position. The second tube and clamp assembly is used to relieve gases and / or pressures generated within the bag during the filling operation. The third connecting tube 89 may be identical to the second tube and clamp assembly and is used as an auxiliary inlet and outlet for recirculation of bag contents.

Similarly, the in-process solution bag 82 has three inlet and / or outlet connecting tubes. The first connecting tube 91 is used as an outlet to pump the solution out of the bag. The second connecting tube 92 serves as a return inlet to allow recirculation of the retentate. The third connecting tube 93 again serves to relieve gases and / or excessive pressures generated within the bag.

The filtrate collection bag and the in-process solution bag are connected to the filtration manifold tube, generally identified as 94 in Fig. 7. Fig. 8 shows the relative positions of the pinch valves 95 and 96 and the position of three pressure sensors 97, 98 and 99. Fig. 9 shows the insertion of the manifold tubes into the peristaltic pump 45.

Before starting the pump 45, all the manual closing clamps are opened, except those that relieve gases and / or pressures generated in the bags. Initially valve 95 is closed and valve 96 is open, while pump 45 begins to recirculate the solution contained in the process solution bag 82 through a tangential flow filtration system 101. The air contained in the tubes and in the tangential flow filtration system 101 ends up in the process solution bag 82, from where it is released to the outside through a sterilizing air filter (not shown). Once the optimum recirculation flow rate of the pump has stabilized, the pinch valve 95 is opened and the filtrate is collected.

Microfiltration or ultrafiltration can be carried out either at constant flow rate or at constant pressure. Appropriate software programs for automating the filtration process through the use of controller 46 are described in US Patents 5,947,689 and US 6,350,382, and US Patent Application publication 2002/0043487.

It will be understood that the embodiments of the present invention that have been described are illustrative of some of the applications of the principles of the present invention. Many modifications can be made by those skilled in the art without departing from the scope of the invention as claimed.

References cited in description

This list of references cited by the applicant serves only to aid the reader and is not part of the European Patent document. Although great care has been taken in compiling the references, errors or omissions cannot be excluded, and the EPO disclaims any responsibility in this regard.

Patent documents cited in the description

US 5480063 A, Keys [0005]

US 5680960 A [0005]

EP 1236644 A [0005]

US 5947689 A [0034] [0036] [0058] [0065]

US 6350382 B [0034] [0036] [0058] [0065]

US 20020043487 A [0034] [0036] [0058].

US 200200434487 A [0065]