Reactor, reactor interior monitoring system, and reactor interior monitoring method

Abstract

To efficiently monitor reactions.SOLUTION: A dialysis unit 3 is provided within a main body of a culture tank 1, which is filled with culture medium. The dialysis unit 3 is equipped with a dialysis membrane 31 that allows substances with molecular weights less than that of proteins to pass through. The dialysis unit 3 is also fluidly connected to a pretreatment unit 4 for pretreating a low-molecular-weight substance M recovered by the dialysis unit 3, a first solvent tank containing a first solvent, and a first liquid delivery unit for delivering the first solvent contained in the first solvent tank to the dialysis unit 3.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a reactor, a reactor in-reactor monitoring system, and a reactor in-reactor monitoring method. [Background technology]

[0002] In the production of biopharmaceuticals and food products, it is important to understand the state of the internal culture environment of a reactor. Deterioration of the internal culture environment of a reactor not only reduces the yield of the desired product, but also affects the quality of the product. To understand the state of the internal culture environment of a reactor, a method is generally used in which the solution inside the reactor is sampled and analyzed to examine the internal culture environment of the reactor (offline analysis). In this case, a wide range of analytical targets are analyzed, including dissolved oxygen concentration, pH, nutrients such as amino acids and sugars, cell and bacterial density, turbidity, product concentration, and metabolites. Appropriate analytical equipment is then used depending on the analytical target.

[0003] On the other hand, methods for analyzing the solution inside a reaction vessel without sampling it are also being considered. Patent Document 1 discloses a method for analyzing the inside of a wine fermentation vessel by Raman spectroscopy (see FIG. 1).

[0004] Patent Document 2 discloses a continuous culture method and apparatus, which "includes a culture step of culturing microorganisms or cells in a culture solution contained in a culture tank 1, and a membrane separation step of separating products in the culture solution produced by the microorganisms or cells using a membrane separation device 3, wherein the membrane separation device 3 is installed in the culture tank 1, and the membrane separation step is a forward osmosis step in which the culture solution in the culture tank is brought into contact with a draw solution having an osmotic pressure higher than that of the culture solution via a separation membrane, thereby causing products produced by the microorganisms or cells in the culture solution to pass through the separation membrane and move into the draw solution" (see abstract).

[0005] Patent Document 3 discloses a liquid chromatograph directly coupled mass spectrometer that "can increase the detection sensitivity in a liquid chromatograph directly coupled mass spectrometer incorporating a sample trapping device" (see abstract).

[0006] Non-Patent Document 1 describes an analytical method using a dialysis cell and a sample loop (see page 231, right column, line 9 to page 232, left column, line 15). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] U.S. Patent Publication No. 2021 / 0255040 [Patent Document 2] Japanese Patent Application Publication No. 2019-146515 [Patent Document 3] Japanese Patent Application Publication No. 8-201343 [Non-patent literature]

[0008] [Non-Patent Document 1] Yasuyuki Kobayashi, Takahisa Yamamoto, "Application of dialysis to ion analysis", Analysis, 2021, pp. 231-235 Summary of the Invention [Problem to be solved by the invention]

[0009] As mentioned above, when analyzing chemical substances in a reaction vessel, it is common to sample a portion of the solution in the reaction vessel and analyze the sample using precision analytical means such as a mass spectrometer (offline analysis). In this case, when the solution is sampled, there is a risk that dirt, bacteria, viruses, etc. may enter the inside of the reaction vessel and contaminate the inside of the reaction vessel. In addition, because of the risk of contamination inside the reaction vessel, there is a problem in that it is difficult to sample the solution frequently and closely examine the changes in the concentration of the components to be analyzed.

[0010] The present invention has been made in view of the above background, and an object of the present invention is to efficiently monitor a reaction. [Means for solving the problem]

[0011] In order to solve the above-mentioned problems, the present invention provides a dialysis section in the main body of a reaction tank filled with a reaction solution, and the dialysis section is provided with a dialysis membrane that allows substances with molecular weights less than that of proteins to pass through. Other solutions will be described as appropriate in the embodiments. [Effects of the Invention]

[0012] According to the present invention, reaction monitoring can be carried out efficiently. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 is a diagram showing the configuration of a reaction tank monitoring system according to a first embodiment. [Figure 2] FIG. 2 is a diagram showing the configuration of a liquid delivery unit of the reaction tank monitoring system in the first embodiment. [Figure 3] FIG. 2 is a diagram showing the configuration of a pre-processing unit of the reaction tank monitoring system in the first embodiment. [Figure 4] FIG. 2 is a functional block diagram of a control device. [Figure 5A] 1 is a flowchart (part 1) showing the operation of the reaction tank monitoring system in the first embodiment. [Figure 5B] 4 is a flowchart (part 2) showing the operation of the reaction tank monitoring system in the first embodiment. [Figure 6] FIG. 1 is a diagram (part 1) showing the operation of a pre-processing unit of the reaction tank monitoring system in the first embodiment. [Figure 7] FIG. 2 is a diagram (part 2) showing the operation of the pre-processing unit of the reaction tank monitoring system in the first embodiment. [Figure 8]FIG. 10 is a diagram (part 3) showing the operation of the pre-processing unit of the reaction tank monitoring system in the first embodiment. [Figure 9] FIG. 4 is a diagram (part 4) showing the operation of the pre-processing unit of the reaction tank monitoring system in the first embodiment. [Figure 10] FIG. 2 is a timing chart showing the steps performed by the reaction tank monitoring system in the first embodiment. [Figure 11] FIG. 10 is a diagram showing the configuration of a reaction tank monitoring system according to a second embodiment. [Figure 12] FIG. 10 is a diagram showing the configuration of a reaction tank monitoring system according to a third embodiment. [Figure 13] FIG. 10 is a diagram showing another example of the reaction tank monitoring system according to the third embodiment. [Figure 14] 10 is a flowchart showing the steps of a method for injecting a liquid culture medium in a reaction tank monitoring system according to a third embodiment. [Figure 15] FIG. 10 is a diagram showing the configuration of a reaction tank monitoring system according to a fourth embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0014] Next, modes for carrying out the present invention (referred to as "embodiments") will be described in detail with reference to the drawings as appropriate.

[0015] [First embodiment] FIG. 1 is a diagram showing the configuration of a reaction tank monitoring system Z according to the first embodiment.

[0016] The reaction tank monitoring system Z is composed of a liquid delivery unit 2, a dialysis unit 3, a pre-treatment unit 4, an analysis unit 5, and piping P for connecting these. Furthermore, the reaction tank monitoring system Z is provided with a control device 6 which is a control unit for controlling the liquid delivery unit 2, the pre-treatment unit 4, and the analysis unit 5.

[0017] The following explanation will be given regarding the reactor monitoring system Z that monitors the internal culture environment of a culture tank 1 for producing biopharmaceuticals, but the same applies to monitoring the inside of a reactor used in food production, etc.

[0018] The user starts culturing in a closed culture environment such as a culture tank 1 by first introducing sugars as nutrients for the cells, a liquid medium L21, and genetically modified cells (hereinafter referred to as cells) as the cells to be cultured into the culture tank 1. The liquid medium L21 contains desired components such as amino acids and salts that adjust the culture environment.

[0019] Specifically, the user installs a sterilized dialysis unit 3 inside (inside the main body of) a culture tank 1, which is a closed-system reaction tank with a sterilized interior. Then, the user puts into the culture tank 1 a culture solution L, which is a reaction solution containing sugar, a liquid medium L21, cells to be cultured, and the like.

[0020] The dialysis unit 3 is provided inside the culture tank 1. The dialysis unit 3 is provided with a dialysis membrane 31. The dialysis membrane 31 allows low-molecular-weight chemical substances (hereinafter referred to as low-molecular-weight substances M) having a molecular weight less than a predetermined molecular weight to pass through, but does not allow substances having a molecular weight equal to or greater than the predetermined molecular weight to pass through. The predetermined molecular weight is the molecular weight of a protein, and the dialysis membrane 31 allows substances having a molecular weight less than that of a protein to pass through. Conversely, the dialysis membrane 31 does not allow substances having a molecular weight equal to or greater than that of a protein to pass through. In other words, the dialysis unit 3 is provided in the culture solution L filled in the culture tank 1.

[0021] The molecular weight of the sample low-molecular-weight substance M is less than the molecular weight of a protein (at least 5000 or more), that is, about several hundred (less than 5000). In other words, the openings of the dialysis membrane 31 are large enough to prevent the passage of substances having molecular weights equal to or greater than the molecular weight of a protein.

[0022] In addition, a solvent is sent from the liquid sending unit 2 to the dialysis unit 3. As a result, the inside of the dialysis unit 3 is filled with the solvent. After a certain period of time has passed in this state, the low molecular weight substances M present inside the culture tank 1 are absorbed into the solvent filling the dialysis unit 3 through the dialysis membrane 31 of the dialysis unit 3. The low molecular weight substances M include components of the liquid medium L21 and substances released from cells due to their metabolism. Eventually, the concentration of the low molecular weight substances contained in the solvent approaches the concentration of the culture solution L filling the inside of the culture tank 1.

[0023] In addition, in this embodiment, the dialysis unit 3 has a cylindrical shape, and the dialysis membrane 31 is provided on the peripheral wall of the dialysis unit 3, but this is not limited to this. For example, the dialysis unit 3 may have a hollow prismatic shape, and the dialysis membrane 31 may be provided on the peripheral wall of the dialysis unit 3. Alternatively, the dialysis unit 3 may have a hollow spherical shape, and the dialysis membrane 31 may be provided on the spherical surface.

[0024] That is, as described above, in a state where the dialysis unit 3 is provided inside the culture tank 1 into which sugar, liquid medium L21, and cells to be cultured have been introduced, the solvent is delivered from the liquid delivery unit 2. As a result, the inside of the dialysis unit 3 is filled with the solvent.

[0025] After the inside of the dialysis unit 3 is filled with the solvent, cells are cultured in the culture tank 1. As the cells are cultured, low molecular weight substances M are released from the cells as metabolites. The released low molecular weight substances M pass through the dialysis membrane 31 due to osmotic pressure and permeate into the inside of the dialysis unit 3. As a result, the low molecular weight substances M are mixed with the solvent inside the dialysis unit 3. The mixture of the solvent and the low molecular weight substances M is called solution L0 (see FIG. 2).

[0026] Next, the solution L0 containing the low molecular weight substance M filled inside the dialysis unit 3 is sent to the pretreatment unit 4 by the solvent sent from the liquid sending unit 2. As a result, the inside of the dialysis unit 3 is filled with new solvent, and the next dialysis can be started in the dialysis unit 3.

[0027] The solution L0 containing the low molecular weight substance M sent to the pretreatment unit 4 undergoes pretreatment such as desalting, concentration, and separation in the pretreatment unit 4, and is then sent to the analysis unit 5. The low molecular weight substance M is then analyzed by the analysis unit 5. In this way, the pretreatment unit 4 is used to pretreat the low molecular weight substance M recovered by the dialysis unit 3. The analysis unit 5 is, for example, a mass spectrometer 51, and is used to analyze the low molecular weight substance M treated in the pretreatment unit 4.

[0028] As described above, the dialysis membrane 31 provided in the dialysis unit 3 has openings that do not allow proteins or larger substances to pass through. Therefore, even if the solution L0 filling the inside of the dialysis unit 3 contains polymers, bacteria, viruses, etc., these cannot pass through the dialysis membrane 31 of the dialysis unit 3. This significantly reduces the risk of the internal culture environment of the culture tank 1 being contaminated by polymers, bacteria, viruses, etc., even if the solution L0 filling the inside of the dialysis unit 3 contains polymers, bacteria, viruses, etc. Furthermore, when providing the dialysis unit 3 inside the culture tank 1, the user thoroughly sterilizes the dialysis unit 3. This reduces the risk of the interior of the culture tank 1 being contaminated by bacteria, etc., when recovering the low-molecular-weight substance M present inside the culture tank 1.

[0029] Generally, when analyzing the internal culture environment of the culture tank 1, the lid of the culture tank 1 is opened and the culture solution L containing bacteria and the like is collected. This carries the risk of contaminating the inside of the culture tank 1.

[0030] In contrast, in the reaction tank monitoring system Z described in this embodiment, the dialysis unit 3 is provided in advance inside the culture tank 1, so the lid of the culture tank 1 is not opened or closed when recovering the low molecular weight substance M. This reduces the risk of contamination due to the opening and closing of the lid of the culture tank 1.

[0031] The analysis targets (low molecular weight substances M) in the analysis unit 5 are nutrients for cell culture, such as sugars and amino acids, and metabolites produced by cellular metabolism. If an abnormality occurs during the cell culture process, the amount of nutrients consumed and the components and amounts of metabolites change. In such a state, the quality of the product (biopharmaceuticals, etc.) produced inside the culture tank 1 decreases. The reaction tank monitoring system Z of the first embodiment can periodically analyze the low molecular weight substances M inside the culture tank 1 without contaminating the inside of the culture tank 1. Therefore, the user can quickly determine if an abnormality has occurred in the culture without having to consider contamination caused by opening and closing the lid of the culture tank 1, etc.

[0032] The solvent sent from the liquid sending unit 2 to the dialysis unit 3 preferably has a composition that does not affect the culture environment even if it penetrates into the culture tank 1 during the dialysis process. Specifically, water, a solution containing salt such as Ringer's solution, liquid medium L21, etc. can be used as the solvent. By performing dialysis for a sufficient period of time, the concentration of the low molecular weight substance M inside the dialysis unit 3 will reflect the concentration of the low molecular weight substance M present inside the culture tank 1.

[0033] Generally, cells are cultured for 2 to 3 days.

[0034] (liquid delivery section 2) 2 is a diagram showing the configuration of the liquid delivery unit 2 of the reaction vessel monitoring system Z in the first embodiment. FIG. 1 will be referred to as appropriate.

[0035] The liquid delivery unit 2 is composed of a first solvent tank 201, a first liquid delivery pump 202 which is a first liquid delivery unit, a first on-off valve 211a which is a first liquid delivery control unit, and a second on-off valve 211b which is a second liquid delivery control unit. The first liquid delivery pump 202, the first on-off valve 211a, and the second on-off valve 211b are controlled by the control device 6. The first solvent tank 201 contains a first solvent L11.

[0036] The first liquid supply pump 202 supplies the first solvent L11 stored in the first solvent tank 201 to the dialysis unit 3. The first solvent tank 201, the first liquid supply pump 202, the first on-off valve 211a, and the second on-off valve 211b are each connected to the dialysis unit 3 via a pipe P so as to be able to flow therethrough. The first on-off valve 211a controls the supply of the first solvent L11 from the first liquid supply pump 202 to the dialysis unit 3. The second on-off valve 211b controls the supply of the solution L0 containing the low-molecular-weight substance M from the dialysis unit 3 to the pretreatment unit 4. Hereinafter, the solution L0 refers to the first solvent L11 mixed with the low-molecular-weight substance M.

[0037] First, the control device 6 opens the first on-off valve 211a and the second on-off valve 211b. Furthermore, the control device 6 operates the first liquid supply pump 202 to supply the first solvent L11 contained in the first solvent tank 201 to the dialysis unit 3. After the inside of the dialysis unit 3 is filled with the first solvent L11, the control device 6 closes the first on-off valve 211a and the second on-off valve 211b and stops the first liquid supply pump 202. After a predetermined time has elapsed in this state, the low molecular weight substance M in the culture tank 1 moves due to osmotic pressure into the first solvent L11 contained in the dialysis unit 3. This allows the dialysis unit 3 to recover the low molecular weight substance M.

[0038] (Preprocessing section 4) 3 is a diagram showing the configuration of the pre-treatment unit 4 of the reaction tank monitoring system Z in the first embodiment. FIG. 1 will be referred to as appropriate.

[0039] The pre-treatment unit 4 is equipped with two six-way valves 401 (a first six-way valve 401a serving as a first flow path switching unit and a second six-way valve 401b serving as a second flow path switching unit). The pre-treatment unit 4 also includes a pre-trap 402 serving as a holding unit, a first waste liquid tank 405a, a second solvent tank 403a, and a second liquid delivery pump 404a serving as a second liquid delivery unit. The pre-treatment unit 4 is also equipped with a concentration column 411 serving as a concentration unit, a third solvent tank 403b, a third liquid delivery pump 404b serving as a third liquid delivery unit, and a second waste liquid tank 405b. The pre-treatment unit 4 is also equipped with a separation column 413 serving as a separation unit, a switching valve 415, and a third waste liquid tank 405c. The first six-way valve 401 a , the second six-way valve 401 b , the second liquid feed pump 404 a , the third liquid feed pump 404 b , and the switching valve 415 are controlled by the control device 6 .

[0040] The first six-way valve 401a is fluidly connected to a pipe P from the dialysis unit 3, a pretrap 402 that temporarily stores the solution L0 to be analyzed, and a first waste liquid tank 405a. Furthermore, the first six-way valve 401a is fluidly connected to a second liquid feed pump 404a, a second solvent tank 403a, and a second six-way valve 401b. The first six-way valve 401a to the analysis unit 5 are fluidly connected by the pipe P. The second six-way valve 401b is connected to a concentration column 411, a third liquid feed pump 404b, a third solvent tank 403b, a second waste liquid tank 405b, a separation column 413, a switching valve 415, a third waste liquid tank 405c, and the analysis unit 5. The second six-way valve 401b to the analysis unit 5 are fluidly connected by the pipe P.

[0041] The pretrap 402 temporarily holds the solution L0 recovered in the dialysis unit 3. The second solvent tank 403a contains the second solvent L12 to be sent to the pretrap 402. The second liquid supply pump 404a sends the second solvent L12 to the pretrap 402. The first six-way valve 401a switches the flow path of the liquid sent to the pretrap 402.

[0042] The concentration column 411 concentrates the low molecular weight substance M in the solution L0 in which the low molecular weight substance M is mixed with the first solvent L11. In the separation column 413, each substance contained in the low molecular weight substance M concentrated in the concentration column 411 is separated. The third solvent tank 403b contains the third solvent L13 that is sent to the separation column 413 via the concentration column 411.

[0043] The third liquid delivery pump 404b delivers the third solvent L13 to the separation column 413 via the concentration column 411. The second six-way valve 401b selects whether the solvent to be delivered to the concentration column 411 is the second solvent L12 or the third solvent L13, and delivers the third solvent L13 to the separation column 413 via the concentration column 411.

[0044] In this embodiment, a configuration using two six-way valves 401 (a first six-way valve 401a and a second six-way valve 401b) is shown, assuming that a mass spectrometer 51 is used in the analysis unit 5. However, if the type of analysis unit 5 is different, a pre-processing unit 4 appropriate for the type can be used. Furthermore, even when a mass spectrometer 51 is used as the analysis unit 5, an appropriate pre-processing unit 4 can be used depending on the measurement target, and the configuration is not limited to that shown in FIG.

[0045] The detailed operation of the pre-processing unit 4 will be described later with reference to FIGS.

[0046] (Control device 6) FIG. 4 is a functional block diagram of the control device 6.

[0047] The control device 6 is configured by a PC (Personal Computer) or the like, and includes at least a calculation device 601 , a storage device 602 , a communication device 603 , and a memory 610 .

[0048] The arithmetic device 601 is composed of a CPU (Central Processing Unit), a GPU (Graphic Processing Unit), etc. The storage device 602 is composed of an HDD (Hard Disk Drive), an SSD (Solid State Drive), etc. The communication device 603 transmits control signals to the first liquid feed pump 202, the second liquid feed pump 404a, the third liquid feed pump 404b, etc. of the reaction vessel monitoring system Z. The communication device 603 also transmits control signals to the first on-off valve 211a, the second on-off valve 211b, the first six-way valve 401a, the second six-way valve 401b, the switching valve 415, etc. The memory 610 is composed of a RAM (Random Access Memory), etc.

[0049] Then, the program stored in the storage device 602 is loaded into the memory 610. Then, the loaded program is executed by the arithmetic device 601, whereby the control unit 611 is realized.

[0050] The control unit 611 controls the operation of the first liquid feed pump 202, the second liquid feed pump 404a, the third liquid feed pump 404b, etc. of the reaction tank monitoring system Z by sending control signals to these. Furthermore, the control unit 611 controls the operation of the first on-off valve 211a, the second on-off valve 211b, the first six-way valve 401a, the second six-way valve 401b, the switching valve 415, etc. In this way, the control device 6 can control the first liquid feed pump 202, the first six-way valve 401a, the second liquid feed pump 404a, the second six-way valve 401b, the third liquid feed pump 404b, etc.

[0051] (Operation of pre-processing unit 4) 5A and 5B are flowcharts showing the operation of the reaction tank monitoring system Z in the first embodiment. Also, FIGS. 6 to 9 are diagrams showing the operation of the pretreatment unit 4 of the reaction tank monitoring system Z in the first embodiment. The operation of the reaction tank monitoring system Z, which is a reaction tank monitoring method, will be described with reference to FIGS. 5A and 5B as well as FIGS. 6 to 9. In the following description of FIGS. 6 to 9, the step numbers are the step numbers of the flowcharts shown in FIGS. 5A and 5B.

[0052] FIG. 6 is a diagram showing the dialysis process.

[0053] First, the control device 6 switches the flow paths of the first six-way valve 401a and the second six-way valve 401b to form the flow paths shown in FIG. 6. Specifically, the control device 6 switches the flow paths of the first six-way valve 401a to allow communication between the first solvent tank 201 and the second on-off valve 211b, the pretrap 402, and the first waste tank 405a. The control device 6 also switches the flow paths of the first six-way valve 401a and the second six-way valve 401b to allow communication between the second solvent tank 403a, the second liquid feed pump 404a, the concentration column 411, and the second waste tank 405b. Furthermore, the control device 6 switches the flow paths of the second six-way valve 401b to allow communication between the third solvent tank 403b, the third liquid feed pump 404b, the separation column 413, the switching valve 415, the third waste tank 405c, and the analysis unit 5.

[0054] Then, the control device 6 opens the first on-off valve 211a and the second on-off valve 211b (S101 in FIG. 5A). Then, the control device 6 sends the first solvent L11 stored in the first solvent tank 201 to the dialysis unit 3 by the first liquid-sending pump 202 (S102). Step S102 is a first liquid-sending step in which the control device 6 introduces the first solvent L11 into the dialysis unit 3 by the first liquid-sending pump 202.

[0055] After the inside of the dialysis unit 3 is filled with the first solvent L11, the control device 6 closes the first on-off valve 211a and the second on-off valve 211b (S103) and stops the first liquid supply pump 202 (S104). The control device 6 determines whether the inside of the dialysis unit 3 is filled with the first solvent L11 based on the liquid supply time by the first liquid supply pump 202, etc. Step S104 is an introduction stop step in which the control device 6 stops the introduction of the first solvent L11 by the first liquid supply pump 202.

[0056] The dialysis unit 3 is filled with the first solvent L11, and thereby the low molecular weight substance M (schematically represented by a triangle or a square in FIGS. 6 to 9) inside the culture tank 1 is dialyzed (S105). As a result, the low molecular weight substance M is recovered in the dialysis unit 3. Step S105 is a dialysis step in which the control device 6 waits for a predetermined time to transfer the low molecular weight substance M from the culture tank 1 to the dialysis unit 3, thereby performing dialysis using the dialysis unit 3.

[0057] Before or during dialysis by the dialysis unit 3, the control device 6 may cause the second solvent L12 stored in the second solvent tank 403a to be sent to the concentration column 411 by the second liquid sending pump 404a. This allows the concentration column 411 to be washed and initialized. The second solvent L12 that has passed through the concentration column 411 is sent to the second waste liquid tank 405b.

[0058] Furthermore, before or during dialysis by the dialysis unit 3, the control device 6 may cause the third liquid supply pump 404b to send the third solvent L13 contained in the third solvent tank 403b (white arrow A2). This also cleans and initializes the separation column 413. At this time, the control device 6 sets the switching valve 415 to the third waste liquid tank 405c side, so that the third solvent L13 that has passed through the separation column 413 is sent to the third waste liquid tank 405c.

[0059] FIG. 7 is a diagram showing the process of transferring the low molecular weight substance M recovered in the dialysis section 3 to the pre-trap 402.

[0060] When the low-molecular-weight substance M is collected by the dialysis unit 3, the control device 6 opens the first on-off valve 211a and the second on-off valve 211b (S111). Furthermore, the control device 6 sends the first solvent L11 stored in the first solvent tank 201 to the dialysis unit 3 using the first liquid-sending pump 202 (S112: outline arrow A1). The control device 6 determines whether the low-molecular-weight substance M has been collected by the dialysis unit 3 based on whether a first predetermined time has elapsed since dialysis by the dialysis unit 3 was started. The first predetermined time is a time sufficient for the low-molecular-weight substance M to be collected by the dialysis unit 3, and is a time preset in the control device 6 by a user or the like. Step S112 is a second liquid-sending step in which the control device 6 sends the first solvent L11 to the dialysis unit 3 using the first liquid-sending pump 202, thereby moving the low-molecular-weight substance M inside the dialysis unit 3 to the pre-trap 402.

[0061] In steps S111 and S112, the solution L0 in which the low-molecular-weight substance M is mixed with the first solvent L11 is pushed toward the pre-trap 402. That is, the low-molecular-weight substance M recovered in the dialysis unit 3 is pushed toward the pre-trap 402. The control device 6 stops the first solution-feed pump 202 when a second predetermined time has elapsed, based on the flow rate and solution-feed time of the first solution-feed pump 202 (S113). The second predetermined time is a time sufficient for all the low-molecular-weight substance M to be discharged from the dialysis unit 3 into the pre-trap 402, and is a time that is preset in the control device 6 by a user or the like.

[0062] At this time, by making the capacity of the pre-trap 402 larger than the capacity of the dialysis unit 3, the low molecular weight substance M recovered in the dialysis unit 3 can be reliably recovered in the pre-trap 402. This allows the low molecular weight substance M recovered in the dialysis unit 3 to be used without waste.

[0063] After the low molecular weight substance M is collected in the pre-trap 402, the control device 6 closes the first on-off valve 211a and the second on-off valve 211b (S114). This allows the dialysis unit 3 to start dialysis for collecting the next low molecular weight substance M. In this way, when the second predetermined time has elapsed since the start of steps S111 and S112, the control device 6 stops the first liquid supply pump 202 and closes the first on-off valve 211a and the second on-off valve 211b.

[0064] FIG. 8 is a diagram showing the process of concentrating the low molecular weight substance M and desalting to remove unnecessary salts.

[0065] After the low-molecular-weight substance M is collected in the pretrap 402, the controller 6 switches the first six-way valve 401a and the second six-way valve 401b to the flow paths shown in Fig. 8 (S121 in Fig. 5B). Specifically, the controller 6 switches the flow path of the first six-way valve 401a so that the first solvent tank 201, the second on-off valve 211b, and the first waste liquid tank 405a can communicate with each other. The controller 6 also switches the flow paths of the first six-way valve 401a and the second six-way valve 401b so that the second solvent tank 403a, the second liquid feed pump 404a, the pretrap 402, the concentration column 411, and the second waste liquid tank 405b can communicate with each other. Furthermore, the control device 6 switches the flow path of the second six-way valve 401b so that the third solvent tank 403b, the third liquid delivery pump 404b, the separation column 413, the switching valve 415, the third waste liquid tank 405c, and the analytical unit 5 can communicate with each other (same as in Figures 6 and 7).

[0066] Step S121 is a first switching step in which the controller 6 switches the flow path in the first six-way valve 401a so that the pretrap 402 and the concentration column 411 can flow through.

[0067] The control device 6 then operates the second liquid feed pump 404a. The second solvent L12 contained in the second solvent tank 403a is fed by the second liquid feed pump 404a (S122), whereby the low molecular weight substance M transferred to the pretrap 402 is sent to the concentration column 411. After a third predetermined time has elapsed since the start of step S122, the control device 6 stops the second liquid feed pump 404a (S123). The third predetermined time is a time sufficient for almost all of the low molecular weight substance M (solution L0) in the pretrap 402 to be sent to the concentration column 411, and is a time that is preset in the control device 6 by a user or the like. In step S122, the control device 6 controls the second liquid feed pump 404a to feed the second solvent L12 to the pretrap 402. That is, step S122 is a third liquid transfer step in which the control device 6 transfers the low molecular weight substance M inside the pretrap 402 to the concentration column 411.

[0068] The state of step S123 is maintained for a fourth predetermined time, whereby the low molecular weight substance M in the solution L0 is concentrated in the concentration column 411 (S124). The fourth predetermined time is a time sufficient for the low molecular weight substance M to be concentrated in the concentration column 411, and is a time that is preset in the control device 6 by a user or the like. Step S124 is a concentration step in which the control device 6 waits for a predetermined time to concentrate the low molecular weight substance M in the concentration column 411.

[0069] By using water or a solvent with a high water content as the second solvent L12, the low molecular weight substance M, which has a low affinity for water, is retained on the stationary phase (silica particles, etc.) of the concentration column 411. On the other hand, the salt that was transferred to the pretrap 402 together with the low molecular weight substance M has a high affinity for water, so it passes through the concentration column 411 and is sent to the second waste liquid tank 405b. This causes the salt in the solution L0 to be desorbed, and the low molecular weight substance M is concentrated. When a mass spectrometer 51 (see FIG. 1) is used as the analysis unit 5, salt may interfere with measurement, so it is advisable to separate the low molecular weight substance M from the salt as described above.

[0070] FIG. 9 is a diagram showing the steps of separating and analyzing a small molecule substance M.

[0071] After the low-molecular-weight substance M is concentrated in the concentration column 411, the control device 6 switches the second six-way valve 401b to the flow path shown in FIG. 9 (S131). Specifically, the control device 6 switches the flow path of the second six-way valve 401b so that the pretrap 402 and the second waste liquid tank 405b can communicate with each other. Furthermore, the control device 6 switches the flow path of the second six-way valve 401b so that the third solvent tank 403b, the third liquid feed pump 404b, the concentration column 411, the separation column 413, the switching valve 415, the third waste liquid tank 405c, and the analysis unit 5 can communicate with each other. The setting of the first six-way valve 401a remains in the state switched in step S121. Whether the low-molecular-weight substance M is concentrated in the concentration column 411 is determined by whether the fourth predetermined time has elapsed since step S123.

[0072] Step S131 is a second flow path switching step in which the controller 6 switches the flow path at the second six-way valve 401b so that the concentration column 411 and the separation column 413 can communicate with each other.

[0073] Then, the control device 6 operates the third liquid feed pump 404b. As a result, the third solvent L13 stored in the third solvent tank 403b is fed to the concentration column 411 and the separation column 413 by the third liquid feed pump 404b (S132). As a result, the low molecular weight substance M (solution L0) concentrated in the concentration column 411 is sent to the separation column 413. Further, by feeding the third solvent L13 to the separation column 413, the individual substances constituting the low molecular weight substance M are separated. Step S132 is a fourth liquid feed step in which the control device 6 feeds the third solvent L13 to the concentration column 411 and the separation column 413 by the third liquid feed pump 404b.

[0074] An organic solvent or a solvent with a low water content is used as the third solvent L13. As a result, the low molecular weight substance M is desorbed from the stationary phase of the concentration column 411 and sent to the separation column 413. In the separation column 413, the individual substances constituting the low molecular weight substance M are separated and sent out in order of the substance with the fastest migration speed. As shown in FIG. 9, the control device 6 switches the switching valve 415 to the flow path of the analysis unit 5, so that the individual substances constituting the low molecular weight substance M separated in the separation column 413 are sent to the analysis unit 5 in order (black arrows). The flow path of the switching valve 415 may be switched at any stage as long as it is before step S132 is performed. Then, the analysis unit 5 analyzes the sent low molecular weight substance M (S141). In step S141, the analysis unit 5 analyzes the individual substances constituting the low molecular weight substance M. Step S141 is an analysis step in which the analysis unit 5 analyzes the low molecular weight substance M sent from the separation column 413.

[0075] The analysis unit 5 may be a mass spectrometer 51 using an electrospray ion source, an ultraviolet absorption detector, or the like.

[0076] The series of steps from dialysis to analysis is completed by the analysis step in Fig. 9. After this, the process returns to the dialysis step in Fig. 6. Then, the step of transferring the low molecular weight substance M to the pre-trap 402 in Fig. 7, the concentration and desalination steps in Fig. 8, and the analysis step in Fig. 9 are carried out in this order.

[0077] (Timing chart) 10 is a timing chart showing the steps performed by the reaction tank monitoring system Z in the first embodiment, with reference to FIGS. 5A and 5B as appropriate.

[0078] As shown in FIG. 10, each step is composed of dialysis by the dialysis unit 3 (S201), concentration by the concentration column 411 (S202), separation by the separation column 413 (S203), and analysis by the analysis unit 5 (S204). Step S201 corresponds to steps S101 to S105 in FIG. 5A and corresponds to the dialysis step. Step S202 corresponds to steps S121 to S124 in FIG. 5B and corresponds to the concentration step. Step S203 corresponds to steps S131 to S132 in FIG. 5B and corresponds to the third liquid transfer step. Step S204 corresponds to step S141 in FIG. 5B and corresponds to the analysis step.

[0079] The reaction tank monitoring system Z according to the first embodiment can reduce the risk of contaminating the inside of the culture tank 1. Therefore, the reaction tank monitoring system Z makes it possible to analyze the low molecular weight substance M inside the culture tank 1 at high frequency. Furthermore, the execution of dialysis (S201) and the delivery of the solution L0 inside the dialysis unit 3 are controlled by opening and closing the first opening and closing valve 211a and the second opening and closing valve 211b.

[0080] Therefore, as shown in Fig. 10, dialysis (S201), concentration (S202), separation (S203), and analysis (S204) can be performed in parallel. In other words, while concentration (S202), separation (S203), and analysis (S204) are being performed, dialysis (S201) for the next analysis is being performed.

[0081] In this way, the reaction vessel monitoring system Z according to the first embodiment enables efficient analysis.

[0082] By repeating these steps, the low molecular weight substance M present inside the culture tank 1 is analyzed periodically, and the user can understand the internal culture environment of the culture tank 1 online from changes in its concentration. The estimated time for each step shown in Figures 6 to 9 is several hours for the dialysis step in Figure 6, and about an hour for the steps in Figures 7 to 9. However, since the next dialysis step is starting even while the steps in Figures 7 to 9 (concentration to analysis) are being performed, analysis results are obtained essentially every time the dialysis takes place.

[0083] When manufacturing biopharmaceuticals, if the internal culture environment of the culture tank 1 deteriorates, problems such as cell division becoming difficult to progress will occur. This will result in a lower yield of pharmaceutical production. Therefore, it is necessary to analyze the culture medium and understand the internal culture environment.

[0084] According to the first embodiment, it is possible to reduce the risk of contaminating the inside of the culture tank 1, and therefore it is possible to analyze the low molecular weight substance M inside the culture tank 1 with high frequency. For example, if a substance that would not be detected when the culture is progressing normally is detected, the user can determine that an abnormality has occurred inside the culture tank 1, stop the reaction, and quickly take measures such as checking the inside of the culture tank 1.

[0085] Furthermore, if the analysis of the low molecular weight substance M reveals that there is a lack of nutrients, the user adds nutrients for the cells to the culture tank 1.

[0086] The technology described in Patent Document 1 also makes it possible to analyze the internal culture environment of the culture vessel 1 without opening or uncovering the culture vessel 1. However, in order to perform measurements using Raman spectroscopy as used in Patent Document 1, the concentration of the low molecular weight substance M in the solution L0 must be at a certain level or higher. In the reaction vessel monitoring system Z shown in this embodiment, the low molecular weight substance M dialyzed in the dialysis unit 3 can be sent to the analysis unit 5. This allows a mass spectrometer 51 to be used as the analysis unit 5, making it possible to analyze even low concentrations of low molecular weight substances M. In the technology described in Patent Document 1, the lid of the culture vessel must be opened in order to analyze low concentrations of low molecular weight substances M.

[0087] Furthermore, the technology described in Patent Document 2 is intended to prevent clogging of the filter, and its role is different from that of the dialysis section 3 shown in the first embodiment.

[0088] As described above, according to the first embodiment, there is little risk of contaminating the inside of the culture tank 1, and therefore it is possible to analyze chemical substances inside the culture tank 1 with high frequency. Therefore, the user can quickly grasp any changes in the internal culture environment of the culture tank 1. Furthermore, by taking appropriate measures against changes in the internal culture environment, the internal reaction of the culture tank 1 can be returned to normal, and the yield and quality of the product in the culture tank 1 can be maintained at a high level. As described above, according to the first embodiment, reaction monitoring can be performed efficiently.

[0089] Furthermore, in the reaction tank monitoring system Z shown in the first embodiment, the dialysis unit 3 is installed inside the culture tank 1, so that the system can be made smaller.

[0090] [Second embodiment] Fig. 11 is a diagram showing the configuration of a reaction tank monitoring system Za according to the second embodiment. Fig. 11 shows the culture tank 1a and the liquid delivery unit 2 of the reaction tank monitoring system Za. The other configurations are the same as those of the first embodiment.

[0091] In the second embodiment, a branch pipe 101 is provided in the culture tank 1a. A dialysis unit 3 is provided inside this branch pipe 101. The operations of the first liquid supply pump 202 and the first and second open / close valves 211a and 211b provided in the piping P are similar to those of the liquid supply unit 2 in the first embodiment. By providing the dialysis unit 3 inside such a branch pipe 101, even if the dialysis unit 3 is contaminated, the influence of contamination on the culture tank 1 can be reduced.

[0092] The low molecular weight substances M recovered in the dialysis unit 3 are limited to substances with a molecular weight that can pass through the dialysis membrane 31. For this reason, the branch pipe 101 is provided with a sampling port 102, which is a collection port for collecting a portion of the culture solution L (the liquid inside the branch pipe 101) for the purpose of analyzing macromolecules such as proteins. By providing the sampling port 102, it becomes possible to analyze the culture solution L. However, the sampling port 102 can be omitted.

[0093] If the inside of the branch pipe 101 from the sampling port 102 is contaminated, the components and amount of the low molecular weight substance M recovered in the dialysis unit 3 will also change. Therefore, the contamination inside the branch pipe 101 can be detected by the reaction tank monitoring system Za shown in the second embodiment.

[0094] 11, a third on-off valve 103a and a fourth on-off valve 103b controlled by the control device 6 are provided at both ends of the branch pipe 101. That is, the third on-off valve 103a and the fourth on-off valve 103b are provided between the culture tank 1a and the branch pipe 101. The control device 6 can close the third on-off valve 103a and the fourth on-off valve 103b. With this configuration, the influence of contamination inside the branch pipe 101 on the culture tank 1a can be minimized. However, the third on-off valve 103a and the fourth on-off valve 103b can be omitted.

[0095] A fifth on-off valve 103c may also be provided in the sampling port 102. The control device 6 can close the fifth on-off valve 103c to prevent contamination of the sampling port 102 from reaching the branch pipe 101.

[0096] [Third embodiment] Fig. 12 is a diagram showing the configuration of a reaction tank monitoring system Zb according to a third embodiment. Fig. 12 shows the culture tank 1b and the liquid delivery unit 2 of the reaction tank monitoring system Zb. The other configurations are the same as those of the first embodiment.

[0097] The reaction tank monitoring system Zb is provided with an injection pipe 111 for injecting a predetermined substance into the culture tank 1b. The reaction tank monitoring system Zb also includes a discharge pipe 112. The discharge pipe 112 is provided with a filter 113 for removing cells. The injection pipe 111 is also connected to a liquid medium tank 701 and a fourth liquid feed pump 702 in a fluid-flowable manner. The liquid medium tank 701 contains a liquid medium L21. The fourth liquid feed pump 702 is controlled by the control device 6, and feeds the liquid medium L21 contained in the liquid medium tank 701 to the culture tank 1b. The control device 6 controls the first liquid feed pump 202, the first on-off valve 211a, and the second on-off valve 211b, as in the first and second embodiments. However, to avoid cluttering the diagram, the control lines from the control device 6 to the first liquid delivery pump 202, the first opening / closing valve 211a, and the second opening / closing valve 211b are omitted from Figure 12 and Figure 13 described below.

[0098] In the reaction tank monitoring system Zb shown in Fig. 12, the control device 6 continuously injects liquid medium L21 into the culture tank 1b through an injection pipe 111. Then, a cell-free liquid L22 containing a product (biopharmaceutical or the like) is discharged from an outlet pipe 112 via a filter 113 that removes cells. This enables continuous culture.

[0099] In the reaction tank monitoring system Zb shown in Fig. 12, a dialysis unit 3 is also provided inside the culture tank 1b. As a result, if the internal culture environment of the culture tank 1b changes, a component that counteracts the change can be added to the culture solution L filled inside the culture tank 1b via the injection pipe 111. As a result, the internal culture environment of the culture tank 1b can be adjusted to a desired environment.

[0100] For example, if the analysis by the analysis unit 5 reveals that there is a nutrient shortage in the internal culture environment of the culture tank 1b, it is possible to add the nutrient shortage to the culture solution L via the injection pipe 111. This makes it possible to maintain a favorable internal culture environment in the culture tank 1b.

[0101] 13 is a diagram showing another example (reaction tank monitoring system Zb1) of the reaction tank monitoring system Zb according to the third embodiment. In FIG. 13, the same components as those in FIG. 12 are denoted by the same reference numerals, and the description thereof will be omitted.

[0102] In the reaction tank monitoring system Zb1 shown in Fig. 13, an inlet pipe 111 and an outlet pipe 112 are provided in the culture tank 1b1, similar to that shown in Fig. 12. The dialysis unit 3 is provided in the outlet pipe 112, which discharges the cell-free solution L22. With this configuration, similar to Fig. 12, components can be added via the inlet pipe 111 in addition to continuous culture, making it possible to maintain a favorable internal culture environment in the culture tank 1b1.

[0103] The mesh size of the filter 113 provided for cell removal has a diameter large enough to allow proteins to pass through. In contrast, the mesh size of the dialysis membrane 31 provided in the dialysis unit 3 is large enough to allow low-molecular-weight substances M having molecular weights of several hundred to pass through, but not proteins. Thus, the mesh size of the filter 113 for cell removal is larger than the mesh size of the dialysis membrane 31 provided in the dialysis unit 3. Therefore, the concentration of the low-molecular-weight substances M recovered inside the dialysis unit 3 is approximately the same as the concentration of the low-molecular-weight substances M recovered inside the dialysis unit 3 when the dialysis unit 3 is provided inside the culture tank 1. In other words, the filter 113 and the dialysis membrane 31 allow the low-molecular-weight substances M to pass through. On the other hand, the filter 113 allows substances having molecular weights larger than those of proteins to pass through, but the dialysis membrane 31 does not allow substances having molecular weights larger than those of proteins to pass through. Therefore, in Figures 12 and 13, the concentration of the low molecular weight substance M recovered inside the dialysis unit 3 is approximately the same as the concentration of the low molecular weight substance M recovered inside the dialysis unit 3 when the dialysis unit 3 is installed inside the culture tank 1.

[0104] By the way, the period of continuous culture is about two weeks.

[0105] (flowchart) 14 is a flowchart showing the steps of the method for injecting the liquid medium L21 in the reaction tank monitoring system Zb, Zb1 in the third embodiment, with reference to FIGS. 1, 4, 12, and 13 as appropriate.

[0106] First, the control device 611 acquires the analysis results of the low molecular weight substance M by the analysis unit 5 (S301).

[0107] Next, the control device 611 determines whether or not injection of the liquid medium L21 is necessary based on the analysis result (S302). In step S302, when the control device 611 determines that nutrients and the like are insufficient based on the analysis result, it determines that injection of the liquid medium L21 is necessary.

[0108] If injection of the liquid medium L21 is not necessary (S302 -> not necessary), the control device 611 returns the process to step S301.

[0109] If injection of the liquid medium L21 is necessary (S302 → necessary), the control device 6 injects the liquid medium L21 into the culture tank 1b via the injection pipe 111 (S303). The liquid medium L21 is contained in the liquid medium tank 701 by controlling the fourth liquid feed pump 702. Thereafter, the control device 611 returns the process to step S301.

[0110] In this way, the control device 6 injects the liquid medium L21, which is a predetermined substance, from the injection tube 111 in accordance with the analysis result by the analysis unit 5.

[0111] 14, the user does not need to monitor the state of the culture tank 1b, and therefore, continuous culture can be carried out for 24 hours, for example.

[0112] The reactor monitoring system Zb shown in FIG. 12 and the reactor monitoring system Zb1 shown in FIG. 13 enable continuous culture.

[0113] [Fourth embodiment] Fig. 15 is a diagram showing the configuration of the reaction tank monitoring system Zc according to the fourth embodiment. Fig. 15 shows the culture tank 1 portion of the reaction tank monitoring system Zc. The other configurations are the same as those of the first embodiment.

[0114] In the reaction tank monitoring system Zc shown in FIG. 15, the culture tank 1 is a culture bag 120 in which a dialysis unit 3 has been previously installed. The dialysis unit 3 is fluidly connected (connectable) to the liquid supply unit 2 and pre-treatment unit 4 shown in FIG. 1 via sterile connectors 122b and 122c, which are kept sterile at the time of connection. The sterile connectors 122b and 122c are connectors in the claims. Then, the user puts culture solution L, genetically modified cells, etc. into the culture bag 120 via the sterile connector 122a and the tube 121. Thereafter, the user connects the liquid supply unit 2 and pre-treatment unit 4 shown in FIG. 1 to the culture bag 120 via the sterile connectors 122b and 122c, and starts culturing.

[0115] When a biopharmaceutical is produced, the reaction system including the culture vessel 1 may be configured with consumables. In such a case, a culture bag 120 shown in FIG. 15 can be used.

[0116] The reaction vessel monitoring systems Z, Za to Zc, and Zb1 shown in this embodiment are intended to be applied to the production of pharmaceuticals using genetically modified cells, but can also be applied to the fermentation of alcoholic beverages, miso, and the like.

[0117] The present invention is not limited to the above-described embodiments and includes various modifications. For example, the above-described embodiments have been described in detail to clearly explain the present invention, and the present invention is not necessarily limited to those having all of the described configurations. Furthermore, it is possible to replace part of the configuration of one embodiment with the configuration of another embodiment, and it is also possible to add the configuration of another embodiment to the configuration of one embodiment. Furthermore, it is possible to add, delete, or replace part of the configuration of each embodiment with other configurations.

[0118] Furthermore, the above-described components, functions, control unit 611, storage device 602, etc. may be partly or entirely implemented in hardware by, for example, designing them as integrated circuits. Furthermore, as shown in Fig. 4, the above-described components, functions, etc. may be implemented in software by a processor such as a CPU interpreting and executing a program that implements each function. Information such as the programs, tables, and files that implement each function can be stored in the memory 610, a recording device such as an SSD, or a recording medium such as an IC (Integrated Circuit) card, an SD (Secure Digital) card, or a DVD (Digital Versatile Disc).

[0119] In addition, in each embodiment, the control lines and information lines shown are those that are considered necessary for explanation, and not all control lines and information lines in the product are necessarily shown. In reality, it can be considered that almost all components are interconnected. [Explanation of symbols]

[0120] 1. Fermenter (reactor) 1a Fermenter (reactor) 1b Culture tank (reaction tank) 1b1 Fermenter (reactor) 2. Liquid delivery section 3 Dialysis Department 4 Pretreatment section 5 Analysis Department 6 Control device (control unit) 51 Mass spectrometer 101 Branch pipe 102 Sampling port (collection port) 103a Third shut-off valve 103b Fourth opening and closing valve 103c Fifth shut-off valve 111 Injection tube 112 Discharge pipe 113 Filters 120 culture bags 122a Sterile Connector 122b Sterile Connector (Connector) 122c Sterile Connector (Connector) 201 First solvent tank 202 First liquid delivery pump (first liquid delivery unit) 211a First opening / closing valve (first liquid supply control unit) 211b Second opening / closing valve (second liquid supply control unit) 401 Six-way valve 401a: First six-way valve (first flow path switching unit) 401b Second six-way valve (second flow path switching unit) 402 Pre-trap (retaining part) 403a Second solvent tank 403b Third Solvent Tank 404a Second liquid delivery pump (second liquid delivery unit) 404b Third liquid delivery pump (third liquid delivery unit) 405a First waste tank 405b Second waste tank 405c Third waste tank 411 Concentration column (concentration section) 413 Separation column (separation section) 415 Switching valve 611 Control Unit 701 Liquid medium tank 702 Fourth Liquid Transfer Pump L culture solution L0 solution L11 First Solvent L12 Second Solvent L13 Third Solvent L21 liquid medium L22 Cell removal solution M: Small molecule substance (sample) Z Reactor monitoring system

Claims

1. A dialysis unit is provided in the main body of the reaction tank filled with the reaction solution, The dialysis unit is provided with a dialysis membrane that allows substances with molecular weights less than that of proteins to pass through. Reaction tank.

2. a dialysis unit provided in a main body of a reaction tank filled with a reaction solution; a pretreatment unit for pretreatment of the sample recovered by the dialysis unit; Equipped with The dialysis unit is provided with a dialysis membrane that allows the sample, which is a substance having a molecular weight less than that of a protein, to pass through. Monitoring system inside the reactor.

3. a first solvent reservoir containing a first solvent; a first solution delivery unit for delivering the first solvent contained in the first solvent tank to the dialysis unit; each of which is fluidly connected to the dialysis unit.

3. The reaction tank monitoring system according to claim 2.

4. a first liquid delivery control unit that controls delivery of the first solvent from the first liquid delivery unit to the dialysis unit; a second fluid transfer control unit that controls the transfer of fluid from the dialysis unit to the pretreatment unit; 4. The reaction tank monitoring system according to claim 3.

5. The pre-treatment unit a holding unit for temporarily holding the sample recovered in the dialysis unit; a second solvent tank containing a second solvent to be sent to the holding section; a second liquid delivery unit for delivering the second solvent to the holding unit; a first flow path switching unit for switching a flow path of the liquid sent to the holding unit; a concentration unit for concentrating the sample in a solution in which the sample is mixed with the first solvent; a separation unit for separating each substance contained in the sample concentrated in the concentration unit; a third solvent tank containing a third solvent to be sent to the separation section via the concentration section; a third solvent delivery unit for delivering the third solvent to the separation unit via the concentration unit; a second flow path switching unit that selects the second solvent or the third solvent as a solvent to be sent to the concentration unit and sends the third solvent to the separation unit via the concentration unit; The reaction tank monitoring system according to claim 3, further comprising:

6. an analysis unit for analyzing the sample treated in the pretreatment unit; The reaction tank monitoring system according to claim 2, further comprising:

7. The analysis unit is a mass spectrometer The reactor internal monitoring system according to claim 6 .

8. a branch pipe is provided in the reaction tank, The dialysis unit is provided inside the branch pipe.

3. The reaction tank monitoring system according to claim 2.

9. The branch pipe is provided with a sampling port for sampling the liquid inside the branch pipe. The reactor monitoring system according to claim 8 .

10. A valve is provided between the reaction vessel and the branch pipe. The reactor monitoring system according to claim 8 .

11. The reaction vessel is provided with an injection tube for injecting a predetermined substance.

3. The reaction tank monitoring system according to claim 2.

12. an analysis unit for analyzing the sample treated in the pretreatment unit; a control unit that acquires an analysis result by the analysis unit, The control unit Injecting a predetermined substance from the injection tube according to the analysis result by the analysis unit. The reactor monitoring system according to claim 11 .

13. the reaction vessel is a culture bag, The culture bag can be connected to the pretreatment unit by a connector.

3. The reaction tank monitoring system according to claim 2.

14. a dialysis unit provided in a main body of a reaction tank filled with a reaction solution; a first solution delivery unit that delivers a first solvent to the dialysis unit; a pretreatment unit for pretreatment of the sample recovered by the dialysis unit; A control unit; In a reaction tank monitoring system comprising: the dialysis unit is provided with a dialysis membrane that allows the sample, which is a substance having a molecular weight less than that of a protein, to pass through; The pre-treatment unit a holding section, a first flow path switching section, a second liquid delivery section, a concentrating section, a second flow path switching section, a separating section, and a third liquid delivery section; the control unit is capable of controlling the first liquid delivery unit, the first flow path switching unit, the second liquid delivery unit, the second flow path switching unit, and the third liquid delivery unit; The control unit a first liquid delivery step of introducing the first solvent into the dialysis unit by the first liquid delivery unit; an introduction stopping step of stopping the introduction of the first solvent by the first liquid delivery unit; a dialysis step of waiting for a predetermined time to move the sample from the reaction tank to the dialysis unit, thereby performing dialysis in the dialysis unit; a second liquid delivery step of delivering the first solvent to the dialysis unit by the first liquid delivery unit, thereby moving the sample inside the dialysis unit to the holding unit; a first switching step of switching the flow path in the first flow path switching unit so that the holding unit and the concentration unit can communicate with each other; a third liquid delivery step of delivering a second solvent to the holding section by the second liquid delivery section, thereby moving the sample inside the holding section to the concentrating section; a concentrating step of concentrating the sample in the concentrating unit by waiting for a predetermined time; a second flow path switching step of switching the flow path in the second flow path switching unit so that the concentration unit and the separation unit can communicate with each other; a fourth liquid sending step of sending a third solvent to the concentration section and the separation section by the third liquid sending section; A monitoring method in a reactor that performs the above.

15. The reactor in-monitoring system comprises: an analysis unit for analyzing the sample treated in the pretreatment unit; The analysis unit an analysis step of analyzing the sample sent from the separation unit; The method for monitoring inside a reactor according to claim 14, characterized in that the following is carried out.

16. The dialysis step is performed while the concentration step, the third liquid delivery step, and the analysis step by the analysis unit are being performed. The method for monitoring inside a reactor according to claim 15.

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