Methods for manufacturing biopharmaceuticals

Abstract

The present invention aims to provide a method for producing biopharmaceuticals that can improve the productivity of biopharmaceuticals. [Solution] In a method for producing a biopharmaceutical, in which each of a plurality of devices is fluidly connected by elastic tubes and a process liquid is sequentially flowed through the plurality of the devices, after at least one of the following steps: (a) anion exchange chromatography step, (b) cation exchange chromatography step, (c) concentration / buffer replacement step, (d) virus removal filter filtration step, or (e) final filtration step, a tube squeezer 20 is used to push the process liquid remaining in the tube 14 toward the next device.

Description

Technical Field

[0005] ,

[0001] The present invention relates to a method for producing a biopharmaceutical, in which each of a plurality of devices is fluidly connected by an elastic tube and a process liquid is sequentially flowed through the plurality of devices.

Background Art

[0002] A method for producing a biopharmaceutical in which each of a plurality of devices is fluidly connected by an elastic tube and a process liquid is sequentially flowed through the plurality of devices is known (see, for example, Patent Document 1). This type of biopharmaceutical is produced in a production system using CHO (Chinese Hamster Ovary) cells or the like as a host. The culture supernatant used at this time contains not only the biopharmaceutical drug substance but also impurities derived from host cells such as proteins and DNA and virus particles. Therefore, in the purification process, a plurality of steps such as protein A chromatography, virus inactivation, anion exchange chromatography, cation exchange chromatography, and virus removal are performed.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] However, the demand for biopharmaceuticals is increasing, and improvement in productivity has been desired in order to manage the quality of biopharmaceuticals and produce an amount commensurate with the demand.

[0005] An object of the present invention is to provide a method for producing a biopharmaceutical that can solve the problems of the above-described conventional techniques and improve the productivity of biopharmaceuticals.

Means for Solving the Problems

[0006] The present invention relates to a method for producing biopharmaceuticals in which each of a plurality of devices is fluidly connected by elastic tubes, and a process liquid is sequentially flowed through the plurality of devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) Virus removal filter filtration process, or (e) Final filtration process, The process is characterized by using a tube squeezer to push the remaining liquid in the tube toward the next apparatus after at least one of the steps.

[0007] Furthermore, the present invention relates to a method for producing a biopharmaceutical, in which each of a plurality of devices is fluidly connected by an elastic tube, and a process liquid is sequentially flowed through the plurality of the devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) virus removal filter filtration process, and (e) Final filtration process, The process is characterized by using a tube squeezer to push the remaining process liquid in the tube toward the next apparatus after each of the steps.

[0008] Furthermore, the present invention relates to a method for producing biopharmaceuticals in which each of a plurality of devices is fluidly connected by elastic tubes, and a process liquid is sequentially flowed through the plurality of the devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) virus removal filter filtration process, and (e) Final filtration process, The device has a tube squeezer that, only after the virus removal filter filtration step, pushes the remaining process liquid in the tube toward the next apparatus.

[0009] In the above case, any one of the multiple devices is a filter, and the tube squeezer may be used to push the remaining process liquid in the tube toward the next device only with respect to the tube between the filter and the next device. Alternatively, the tube may be squeezed with the tube squeezer to push the remaining process liquid in the tube toward the next device without removing both ends of the tube from the device. [Effects of the Invention]

[0010] The present invention provides a method for producing biopharmaceuticals that can improve the productivity of biopharmaceuticals. [Brief explanation of the drawing]

[0011] [Figure 1] A schematic diagram of a part of the manufacturing process connected to a chromatography system is shown. [Figure 2] A schematic diagram shows a portion of the manufacturing process to which the UF device is connected. [Figure 3] A schematic diagram shows a portion of the manufacturing process to which a VF (Variable Frequency) device is connected. [Figure 4] A perspective view of the tube squeezer in its open state is shown. [Figure 5] A perspective view of the tube squeezer in its closed state is shown. [Figure 6] A flowchart of the manufacturing process is shown. [Modes for carrying out the invention]

[0012] (First Embodiment) The following describes a method for producing a biopharmaceutical according to one embodiment of the present invention, with reference to the drawings. FIG. 1 shows a schematic diagram of a part of a manufacturing process to which a chromatography system is connected, FIG. 2 shows a schematic diagram of a part of a manufacturing process to which a UF device is connected, FIG. 3 shows a schematic diagram of a part of a manufacturing process to which a VF device is connected, FIG. 4 shows a perspective view of an open tube restrictor, FIG. 5 shows a perspective view of a closed tube restrictor, and FIG. 6 shows a flowchart of a manufacturing process.

[0013] In the manufacturing process of the biopharmaceutical according to this embodiment, a culture broth clarification process, a protein A chromatography process, a virus inactivation process, a depth filter filtration process, an AEX chromatography process (anion exchange chromatography process), a CEX chromatography process (cation exchange chromatography process), an ultrafiltration / diafiltration process, a virus removal filter filtration process, and a final filtration process are performed.

[0014] When performing the AEX chromatography process or the CEX chromatography process among these processes, as shown in FIG. 1, the first buffer tank 1, the chromatography system 2, the second buffer tank 3, the membrane filter 4, and the third buffer tank 5 are fluidly connected.

[0015] The first buffer tank 1 and the chromatography system 2 are fluidly connected by a first tube 11, the chromatography system 2 and the second buffer tank 3 are fluidly connected by a second tube 12, the second buffer tank 3 and the membrane filter 4 are fluidly connected by a third tube 13, and the membrane filter 4 and the third buffer tank 5 are fluidly connected by a fourth tube 14.

[0016] When performing the ultrafiltration / diafiltration process, as shown in FIG. 2, the first buffer tank 1, the UF device 6, the second buffer tank 3, the membrane filter 4, and the third buffer tank 5 are fluidly connected.

[0017] The first buffer tank 1 and the UF device 6 are fluidically connected by the first tube 11, the UF device 6 and the second buffer tank 3 are fluidically connected by the second tube 12, the second buffer tank 3 and the membrane filter 4 are fluidically connected by the third tube 13, and the membrane filter 4 and the third buffer tank 5 are fluidly connected by the fourth tube 14.

[0018] During the virus removal filter filtration process and the final filtration process, the first buffer tank 1, the VF (Virus Free) device 7, the second buffer tank 3, the membrane filter 4, and the third buffer tank 5 are fluidly connected, as shown in Figure 3.

[0019] The first buffer tank 1 and the VF device 7 are fluidically connected by the first tube 11, the VF device 7 and the second buffer tank 3 are fluidically connected by the second tube 12, the second buffer tank 3 and the membrane filter 4 are fluidically connected by the third tube 13, and the membrane filter 4 and the third buffer tank 5 are fluidly connected by the fourth tube 14.

[0020] The process liquid in the first buffer tank 1, which is the final filtration buffer tank for the virus filtration process, flows through the second buffer tank 3, the membrane filter 4, and the third buffer tank 5 in that order after the virus has been removed by the VF device 7.

[0021] Furthermore, each of the devices consisting of the first buffer tank 1, the chromatography system 2, the UF apparatus 6, the second buffer tank 3, the membrane filter 4, and the third buffer tank 5, as well as the first tube 11, the second tube 12, the third tube 13, and the fourth tube 14, may be cleaned after each process and reused in the next process, or they may be replaced each time.

[0022] The first buffer tank 1, the second buffer tank 3, and the third buffer tank 5 are of a type in which resin bags are placed inside stainless steel containers, and each may be the same size or different in size for each process.

[0023] Membrane filter 4 is an MF (Micro Filtration) filter that captures bacteria such as bioburden but allows antibodies to pass through.

[0024] Chromatography System 2 is a purification system that uses chromatography to separate antibodies from impurities.

[0025] The UF device 6 is a filter that uses an ultrafiltration membrane.

[0026] The VF device 7 has a virus removal filter 8 such as Planova (registered trademark). The VF device 7 has a lower section 7a and an upper section 7b, and the downstream end of the lower section 7a and the upstream end of the upper section 7b are fluidly connected via a virus removal filter 8 installed in between, and a storage section 9 is connected to one end of this virus removal filter 8.

[0027] The upstream end of the lower section 7a of the device is connected to the first buffer tank 1 via a tube 11. The downstream end of the upper section 7b of the device is connected to the second buffer tank 3 via a tube 12, and a waste tank 10 is connected in parallel with the second buffer tank 3.

[0028] Each tube 11, 12, 13, and 14 is elastic and, after each process, is squeezed using the tube press 20 to push out any remaining process liquid from tubes 11, 12, 13, and 14 toward the next process. When pushing out the process liquid remaining in tube 12 toward the second buffer tank 3 using the tube press 20, first, the tube 12 is closed by clamping it at position C, which is near the end of tube 12 on the VF device 7 side. Next, tube 12 is removed from the VF device 7 and transported from the room where the VF device 7 is located to the purification room, where it is pushed out from position C toward the second buffer tank 3 using the tube press 20.

[0029] The tube squeezer 20 according to this embodiment is capable of squeezing and crushing the elastic tubes 11, 12, 13, and 14, while simultaneously pushing out the remaining process liquid inside the tubes 11, 12, 13, and 14 toward the next process. As shown in Figure 4, the tube squeezer 20 comprises a first metal frame 22 and a second metal frame 23 that rotate relative to each other around an axis 21.

[0030] The first frame 22 is equipped with a cylindrical metal first roller 24. A handle 25 is integrally attached to the end of the first roller 24, and it is rotated by an operator turning the handle 25. The handle 25 is large enough to apply a large torque to the first roller 24, and the length from the point of force application to the point of application is longer than the distance between the first roller 24 and the shaft 21. Multiple grooves extending in the axial direction are arranged along the outer circumference of the first roller 24, and the first roller 24 has a gear-like cross-section, which allows it to firmly bite into the conveyed tubes 11, 12, 13, and 14, thereby acting as an anti-slip device.

[0031] Furthermore, the first frame 22 is provided with a first gripping portion 26 at one end for the operator to grasp, and a holding mechanism 27 at the other end for holding the shaft 21. This holding mechanism 27 is rotatably attached at one end to the main body of the first frame 22, and the other end is screwed in with a wing bolt 28. The shaft 21 of the second frame 23 is sandwiched and fixed in a groove formed between the one end and the other end of the holding mechanism 27.

[0032] The second frame 23 is equipped with a cylindrical metal second roller 29. This second roller 29 is rotatably mounted and rotates in accordance with the transport of the tubes 11, 12, 13, and 14 sandwiched between it and the first roller 24 of the first frame 22. Multiple grooves extending in the axial direction are arranged along the outer circumference of the second roller 29, and the second roller 29 has a gear-like cross-section, which allows it to firmly grip the transported tubes 11, 12, 13, and 14, thereby acting as an anti-slip mechanism.

[0033] The second frame 23 has a second gripping section 30 at one end for the operator to grip, and a shaft 21 at the other end. A cylindrical guide 31, which is constricted in the middle, is fixedly attached to the shaft 21. The guide 31 is designed to guide the tubes 11, 12, 13, and 14 toward the center of the first roller 24 and the second roller 29, that is, the axial midpoint between the first roller 24 and the second roller 29.

[0034] As shown in Figure 5, the tube squeezer 20 is configured such that, when closed, the first roller 24 and the second roller 29 of the first frame 22 face each other. Furthermore, the axes of the first roller 24 and the second roller 29 are positioned parallel to the axis of the shaft 21.

[0035] The following describes the procedure for using the tube squeezer 20. It is recommended that two workers perform the operation of the tube squeezer 20. When squeezing tubes 11, 12, 13, and 14 to push the process fluid inside them to the next devices 1, 2, 3, 4, 5, and 6, the operator first places tubes 11, 12, 13, and 14 between the separated first frame 22 and second frame 23. The tube squeezer 20 can be separated into the first frame 22 and second frame 23 by releasing the holding mechanism 27. This allows the tube squeezer 20 to hold tubes 11, 12, 13, and 14 without removing both ends of tubes 11, 12, 13, and 14 from the devices 1, 2, 3, 4, 5, and 6.

[0036] Once the tubes 11, 12, 13, and 14 are placed between the first frame 22 and the second frame 23, the worker places the shaft 21 of the second frame 23 between the main body of the first frame 22 and the holding mechanism 27, and screws in the wing bolt 28 to fix the shaft 21 to the holding mechanism 27.

[0037] Once the wing bolt 28 is screwed in to fix the shaft 21 to the holding mechanism 27, the worker holds the tubes 11, 12, 13, and 14 by hand so that they are positioned in the center between the first roller 24 of the first frame 22 and the second roller 29 of the second frame 23, and rotates the second gripping part 30 of the second frame 23 toward the first gripping part 26 of the first frame 22, as shown in Figure 5. As a result, the tubes 11, 12, 13, and 14 are squeezed and crushed between the first roller 24 of the first frame 22 and the second roller 29 of the second frame 23.

[0038] When tubes 11, 12, 13, and 14 are placed between the first roller 24 of the first frame 22 and the second roller 29 of the second frame 23, the operator rotates the handle 25 by hand while pressing the constricted part of the guide 31 against the tubes 11, 12, 13, and 14. This conveys the tubes 11, 12, 13, and 14 to the tube squeezer 20, causing the tube squeezer 20 to move toward the next device, and the process liquid inside the tubes 11, 12, 13, and 14 is pushed toward the next device. In this case, when squeezing tubes 11, 12, 13, and 14 which are made up of multiple tubes joined together, it is desirable not to squeeze the joints between the tubes from a strength standpoint, but to squeeze from a position beyond the joints, that is, only before and after the joints.

[0039] The following describes the manufacturing procedure for producing a monoclonal antibody called MabA in the purification flow, and the verification of improved monoclonal antibody yield, with reference to Figure 6. Note that in this embodiment, the increase in the process solution is calculated to verify the improvement in monoclonal antibody yield; however, in a normal manufacturing procedure, it is not necessary to calculate the increase in the process solution.

[0040] In the manufacturing method according to this embodiment, as shown in Figure 6, the following steps were performed: culture medium clarification step S1, protein A chromatography step S2, virus inactivation step S3, depth filter filtration step S4, AEX chromatography step S5, CEX chromatography step S6, ultrafiltration / dialysis filtration step S7, virus removal filter step S8, and final filtration step S9.

[0041] First, the CHO cell culture medium that produced MabA was filtered to remove the cells and clarified to obtain the cell culture supernatant (Step S1: Culture medium clarification step).

[0042] In step S1, the cell culture supernatant was clarified. Next, the cell culture supernatant was purified by protein A chromatography (step S2: protein A chromatography step) to obtain a protein A chromatography pool containing MabA.

[0043] In step S2, the cell culture supernatant was purified by protein A chromatography. Next, acid was added to the protein A chromatography pool to inactivate the virus (step S3: virus inactivation step). After that, the protein A chromatography pool was neutralized.

[0044] In step S3 and the associated processes, the protein A chromatography pool solution was neutralized. Next, the neutralized protein A chromatography pool solution was loaded onto a depth filter, and the neutralized protein A chromatography pool solution was pushed out with washing buffer to recover MabA. Subsequently, MabA was filtered through membrane filter 4 (step S4: depth filter filtration step) and stored as the depth filter filtered pool solution.

[0045] In step S4, MabA was filtered through the membrane filter 4. Next, the depth filter filtration pool liquid was adjusted to neutral to become the AEX material load liquid, and MabA was purified by AEX chromatography (step S5: AEX chromatography step). The process liquid recovered in this step was adjusted to acidity, filtered through the membrane filter 4, and stored as the AEX material pool liquid. At this time, after recovering the AEX material pool liquid by the usual method, the process liquid in the tube 14 connecting the 0.2 μm membrane filter 4 and the third buffer tank 5, which is the recovery container for the AEX material pool liquid, was squeezed out and recovered using a tube squeezer 20. The increase in the process liquid due to tube squeezing in the AEX chromatography step was calculated from the weight of the AEX material pool liquid before and after tube squeezing.

[0046] In step S5, MabA was purified by AEX chromatography, and then MabA was purified by CEX chromatography, and the CEX material pool solution was recovered (step S6: CEX chromatography step). At this time, after recovering the CEX material pool solution by the usual method, the process solution in the tube 14 connecting the 0.2 μm membrane filter 4 and the third buffer tank 5, which is the recovery container for the CEX material pool solution, was further squeezed out and recovered using a tube squeezer 20, and the increase in the process solution due to tube squeezing in the CEX chromatography step was calculated from the weight of the CEX material pool solution before and after tube squeezing.

[0047] In step S6, MabA was purified by CEX chromatography. Next, the CEX material pool solution was concentrated and buffer-exchanged using an ultrafiltration membrane (concentration / buffer exchange step), filtered through membrane filter 4 to obtain the UF material pool solution (step S7: ultrafiltration / dialysis filtration step). At this time, after recovering the UF material pool solution by the usual method, the process solution in the fourth tube 14 connecting the 0.2 μm membrane filter 4 and the third buffer tank 5, which is the recovery container for the UF material pool solution, was further squeezed out and recovered using a tube squeezer 20. The increase in process solution due to tube squeezing in the ultrafiltration / dialysis filtration step was calculated from the weight of the UF material pool solution before and after tube squeezing.

[0048] In step S7, an ultrafiltration / dialysis filtration process is performed. Next, the UF material pool liquid is filtered through a virus removal filter to remove MabA and obtain the MF material pool liquid (step S8: virus removal filter filtration process).

[0049] In step S8, a filtration process is performed using a virus removal filter. Next, the MF material pool liquid is recovered using the usual method (step S9: final filtration process). Then, the process liquid in the fourth tube 14, which connects the 0.2 μm membrane filter 4 and the third buffer tank 5, which is the recovery container for the MF material pool liquid, is squeezed out and recovered using a tube squeezer 20. The increase in the process liquid due to tube squeezing in the final filtration process is calculated from the weight of the MF material pool liquid before and after tube squeezing.

[0050] According to the above process, the increase in the amount of process liquid due to using the tube squeezing device 20 is shown in Table 1.

[0051] [Table 1]

[0052] As can be seen from this table, by using the tube squeezing device 20, the increase in MabA calculated from the increase in process solution during the AEX chromatography process, CEX chromatography process, concentration / buffer replacement process, and final filtration process was found to be 32-48g, with an average of 40g per lot.

[0053] In the biopharmaceutical manufacturing method according to this embodiment, the fourth tube 14 is squeezed out of the process liquid remaining in the fourth tube 14 using a tube squeezer 20 after at least one of the following steps: (a) anion exchange chromatography, (b) cation exchange chromatography, (c) concentration / buffer replacement, (d) virus removal filter filtration, or (e) final filtration. This allows the process liquid remaining in the fourth tube 14 to be recovered, thereby improving the productivity of the biopharmaceutical.

[0054] Furthermore, it is conceivable to shorten each tube 11, 12, 13, and 14 in order to improve the yield of monoclonal antibodies, but in this case, the degree of freedom in arranging each device would decrease. However, the method for producing biopharmaceuticals according to this embodiment can improve the yield of monoclonal antibodies even if each tube 11, 12, 13, and 14 is kept long, without reducing the degree of freedom in arranging each device.

[0055] Furthermore, because the amount of process fluid that is discarded can be reduced, the amount of waste fluid produced by the manufacturing plant can also be reduced.

[0056] (Second Embodiment) The second embodiment will be described using the same reference numerals for substantially the same components as in the first embodiment.

[0057] In the manufacturing method according to the second embodiment, the only difference from the manufacturing method according to the first embodiment is the step of using the tube squeezing device 20.

[0058] The following describes the manufacturing procedure for producing a monoclonal antibody called MabA in the purification flow, and the verification of improved monoclonal antibody yield, with reference to Figure 5. Note that in this embodiment, the increase in the process solution is calculated to verify the improvement in monoclonal antibody yield; however, in a normal manufacturing procedure, it is not necessary to calculate the increase in the process solution.

[0059] In the manufacturing method according to this embodiment, as shown in Figure 5, the following steps were performed: culture medium clarification step S1, protein A chromatography step S2, virus inactivation step S3, depth filter filtration step S4, AEX chromatography step S5, CEX chromatography step S6, ultrafiltration / dialysis filtration step S7, virus removal filter filtration step S8, and final filtration step S9.

[0060] First, the CHO cell culture medium that produced MabA was filtered to remove the cells and clarified to obtain the cell culture supernatant (Step S1: Culture medium clarification step).

[0061] In step S1, the cell culture supernatant was clarified, and then the cell culture supernatant was purified by protein A chromatography (step S2: protein A chromatography step). At this time, the absorbance at 280 nm was monitored, and the protein A chromatography pool solution, which is the fraction containing MabA, was obtained.

[0062] In step S2, the cell culture supernatant was purified by protein A chromatography. Next, acid was added to the protein A chromatography pool to adjust the pH to 3-4, and the virus was inactivated for 1 hour (step S3: virus inactivation step). After that, the protein A chromatography pool was neutralized to pH 4-6.

[0063] In step S3 and the associated processes, the protein A chromatography pool solution was neutralized. Next, the neutralized protein A chromatography pool solution was loaded onto a depth filter, and the neutralized protein A chromatography pool solution was pushed out with washing buffer to recover MabA. Subsequently, MabA was filtered through membrane filter 4 (step S4: depth filter filtration step) and stored as the depth filter filtered pool solution.

[0064] In step S4, MabA was filtered through the membrane filter 4. Next, the depth filter filtration pool solution was adjusted to neutral to become the AEX material load solution, and MabA was purified by AEX chromatography (step S5: AEX chromatography step). The process solution recovered in this step was adjusted to pH 4-6 using acetic acid, then filtered through a 0.2 μm membrane filter 4 and stored as the AEX material pool solution. At this time, after recovering the AEX material pool solution by the usual method, the process solution in the tube 14 connecting the 0.2 μm membrane filter 4 and the third buffer tank 5, which is the recovery container for the AEX material pool solution, was further squeezed out and recovered using a tube squeezer 20.

[0065] In step S5, MabA was purified by AEX chromatography, and then MabA was purified again by CEX chromatography, and the CEX material pool was recovered (step S6: CEX chromatography step).

[0066] In step S6, MabA is purified by CEX chromatography. Next, the CEX material pool is concentrated and buffer-exchanged using an ultrafiltration membrane (concentration / buffer exchange step), filtered through membrane filter 4, and obtained as the UF material pool (step S7: ultrafiltration / dialysis filtration step).

[0067] In step S7, an ultrafiltration / dialysis filtration process is performed. Next, the UF material pool liquid is filtered through a virus removal filter to remove MabA and obtain the MF material pool liquid (step S8: virus removal filter filtration process).

[0068] In step S8, the filtration process was performed using a virus removal filter. Next, the VF (Virus Free) material pool liquid was recovered using the normal method (step S9: final filtration process). Then, the process liquid in the fourth tube 14 connecting the VF device and the third buffer tank 5, which is the recovery container for the VF material pool liquid, was recovered using a tube squeezer 20. The increase in process liquid due to tube squeezing in the virus removal filter filtration process was calculated from the weight of the VF material pool liquid before and after tube squeezing. The tube squeezer 20 can squeeze with strong force using the lever principle, so even thick-walled and wide tubes for VF material can be squeezed due to the internal pressure. Next, the VF material pool liquid was filtered with a 0.2 μm membrane filter to obtain the virus-filtered pool liquid after filtration.

[0069] According to the above process, the increase in the amount of process liquid due to using the tube squeezing device 20 is shown in Table 2.

[0070] [Table 2]

[0071] As can be seen from this table, by using the tube squeezing device 20, the increase in MabA calculated from the increase in the process liquid during the virus removal filter filtration process was 24-29g, with an average of 26g per lot.

[0072] The biopharmaceutical manufacturing method according to this embodiment comprises (a) anion exchange chromatography step, (b) cation exchange chromatography step, (c) concentration / buffer replacement step, (d) virus removal filter filtration step, and (e) final filtration step. Only after the virus removal filter filtration step, the process liquid remaining in the fourth tube 14 is pushed out towards the next apparatus using a tube squeezer 20. This allows the process liquid remaining in the fourth tube 14 to be recovered, thereby improving the productivity of the biopharmaceutical.

[0073] Furthermore, it is conceivable to shorten each tube 11, 12, 13, and 14 in order to improve the yield of monoclonal antibodies, but in this case, the degree of freedom in arranging each device would decrease. However, the method for producing biopharmaceuticals according to this embodiment can improve the yield of monoclonal antibodies even if each tube 11, 12, 13, and 14 is kept long, without reducing the degree of freedom in arranging each device.

[0074] Furthermore, because the amount of process fluid that is discarded can be reduced, the amount of waste fluid produced by the manufacturing plant can also be reduced.

[0075] The present invention has been described above based on embodiments, but the present invention is not limited thereto. For example, in the above embodiments, the process liquid remaining only in the fourth tube 14 is pushed out toward the next device using the tube squeezer 20, but the invention is not limited thereto. The process liquid remaining in the first tube 11, second tube 12, third tube 13, and membrane filter 4, etc., may also be pushed out toward the next device using the tube squeezer 20. In particular, downstream of the membrane filter 4, the sterility of the process liquid cannot be maintained if the fourth tube 14 is removed, so compressed air cannot be used to push out the process liquid remaining in the fourth tube 14, and it cannot be pushed out even if a peristaltic pump is used, so using the tube squeezer 20 is effective. [Explanation of symbols]

[0076] 1…First buffer tank 2…Chromatography system 3…Second buffer tank 4…Membrane filter 5…Third buffer tank 6…UF device 7...VF device 8…Virus removal filter 9... Storage section 10…Disposal tanks 11…First tube 12...Second tube 13…Third tube 14…Fourth tube 20... Tube squeezer 21...axis 22...First frame 23...2nd frame 24…1st Laura 25... Handle 26...First grip part 27...Retention mechanism 28... Wing bolt 29... Second Laura 30…Second gripping part 31… Guide

Claims

1. In a method for manufacturing biopharmaceuticals, in which each of a plurality of devices is fluidly connected by elastic tubes, and a process liquid is sequentially flowed through the plurality of said devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) Virus removal filter filtration process, or (e) Final filtration process, A method for producing a biopharmaceutical, characterized in that, after at least one of the steps, the process liquid remaining in the tube is squeezed out towards the next apparatus using a tube squeezer.

2. In a method for manufacturing biopharmaceuticals, in which each of a plurality of devices is fluidly connected by elastic tubes, and a process liquid is sequentially flowed through the plurality of said devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) Virus removal filter filtration process, and (e) Final filtration process, A method for producing a biopharmaceutical, characterized in that, after each step, the process liquid remaining in the tube is squeezed out towards the next apparatus using a tube squeezer.

3. In a method for manufacturing biopharmaceuticals, in which each of a plurality of devices is fluidly connected by elastic tubes, and a process liquid is sequentially flowed through the plurality of said devices, (a) Anion exchange chromatography process, (b) Cation exchange chromatography process, (c) Concentration / buffer replacement step, (d) Virus removal filter filtration process, and (e) Final filtration process, A method for producing a biopharmaceutical, comprising having a virus removal filter filtration step, and characterized in that, only after the virus removal filter filtration step, the process liquid remaining in the tube is squeezed out toward the next apparatus using a tube squeezer.

4. A method for producing a biopharmaceutical according to any one of claims 1 to 3, A method for producing a biopharmaceutical, characterized in that one of the plurality of aforementioned devices is a filter, and the tube squeezing device is used to push the process liquid remaining in the tube toward the next aforementioned device only with respect to the tube between the filter and the next aforementioned device.

5. A method for producing a biopharmaceutical according to any one of claims 1 to 3, A method for producing a biopharmaceutical, characterized in that, without removing both ends of the tube from the apparatus, the tube is squeezed with the tube squeezer and the process liquid remaining inside the tube is pushed out toward the next apparatus.

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