TWI841625B - Simulated moving bed chromatographic separation method and simulated moving bed chromatographic separation system - Google Patents

Abstract

A simulated moving bed chromatographic separation method which uses two or more eluents and a circulation system having a plurality of unit packed columns each packed with adsorbent connected endlessly in series via piping to separate, from a stock solution, a weakly adsorbed component, a strongly adsorbed component, and a moderately adsorbed component having adsorption to the adsorbent that is intermediate relative to the other components, wherein in the piping of the circulation system are provided a stock solution supply port F, two or more eluent supply ports D corresponding with the two or more eluents, a discharge port A for a weakly adsorbed fraction containing the weakly adsorbed component, a discharge port B for a moderately adsorbed fraction containing the moderately adsorbed component, and a discharge port C for a strongly adsorbed fraction containing the strongly adsorbed component, and the positions of the stock solution supply port F, the discharge port A, the discharge port B and the discharge port C are in a specific relationship. Also provided is a chromatographic separation system suited to the implementation of this chromatographic separation method.

Description

Simulated mobile layer-by-layer analysis and separation method and simulated mobile layer-by-layer analysis and separation system

The present invention relates to a simulated mobile layer-by-layer analysis and separation method and a simulated mobile layer-by-layer analysis and separation system.

In the chromatographic separation simulating the moving bed method, a plurality of unit packed towers (hereinafter also referred to as "packed towers" and sometimes "columns") filled with an adsorbent having a selective adsorption capacity for a specific component among two or more components contained in a stock solution are connected in series by piping, and the packed tower at the most downstream position is connected to the packed tower at the most upstream position to form an endless circulation system. The circulation system is supplied with raw liquid and elution liquid, and the faster moving part (weakly adsorbent part) and slower moving part (strongly adsorbent part) in the circulation system are simultaneously extracted from different positions, and the part with a medium moving speed (medium adsorbent part) is extracted as needed. Then, the raw liquid supply position, the elution liquid supply position, the extraction position of the weakly adsorbent part, the extraction position of the medium adsorbent part, and the extraction position of the strongly adsorbent part are moved in the fluid circulation direction of the circulation system while maintaining a certain positional relationship. By repeating this operation, the processing operation of the moving layer that can continuously supply the raw liquid is realized in a simulated manner. Patent document 1 discloses a method for continuously separating three or more parts having different affinities with respect to an adsorbent by repeatedly performing the steps of supplying an elution liquid and a stock solution while extracting a medium-adsorbable part, and supplying an elution liquid while extracting a weakly-adsorbable part and a strongly-adsorbable part.

In the conventional chromatographic separation using the simulated moving layer method, represented by the technology described in Patent Document 1, basically one type of solvent is used. Therefore, when a stock solution containing components with strong adsorption to the adsorbent or a stock solution containing components that are prone to tailing (phenomenon of broadening of concentration distribution) is supplied to a circulation system, a large amount of solvent is required to desorb (detach) these components. The use of a large amount of solvent will increase the concentration cost of the extract and also reduce the production of the target purified product per unit of adsorbent.

On the other hand, in the stratification separation of the simulated moving layer method, two or more solvents are also used. For example, Patent Document 2 describes the use of a first solvent with a weaker desorption force and a second solvent with a stronger desorption force, and setting the supply timing of these solvents and the stock solution and the extraction timing of the weakly adsorbable part, the medium adsorbable part, and the strongly adsorbable part to a specific combination, thereby achieving a higher separation efficiency with a smaller amount of adsorbent. [Prior art] [Document] [Patent document]

[Patent document 1] Japanese Patent No. 1998860 [Patent document 2] Japanese Patent No. 4606092

[The problem the invention is trying to solve]

The simulated moving-layer separation method can obtain the target purified object continuously and with high purity, so its applicability in the medical field is also examined. For example, in the production of antibody drugs, in addition to the target antibody, the extract or culture medium of the cultured cells that produce the antibody will also produce fragments that cannot fully function as antibodies due to antibody cleavage, or giant aggregates of antibodies. Generally speaking, the above-mentioned fragments have fewer sites for interaction with the adsorbent and have weaker adsorption to the adsorbent. On the contrary, the adsorption of aggregates to the adsorbent is stronger. Therefore, when applying the simulated shift layer method of analytical separation to the purification of antibody drugs, the target antibody must be extracted as a medium-adsorbable portion showing medium adsorption relative to the adsorbent. On the other hand, both the weakly adsorbable portion and the strongly adsorbable portion must be fully removed at a high removal rate. In addition, in the practical application of such analytical separation, it is also very important to reduce the necessary amount of adsorbent as much as possible and improve the separation efficiency to achieve the purpose of low cost. However, the inventors of the present invention have reviewed the previous simulated shift layer method of analytical separation, represented by the technologies described in the above-mentioned patent documents, and found that it is quite difficult to fully achieve the above-mentioned purpose.

Therefore, the purpose of the present invention is to provide a chromatographic separation method using a simulated moving layer method, which can extract the purified component in the stock solution with high purity using a smaller amount of adsorbent. In addition, the purpose of the present invention is to provide a chromatographic separation system suitable for implementing the above-mentioned chromatographic separation method. [Means for solving the problem]

In view of the above problems, the inventors of the present invention have continuously focused on research and found that in the stratification separation method using the simulated moving layer method, by using two or more solvents and setting the raw liquid supply port, the strong adsorption part extraction port, the medium adsorption part extraction port and the weak adsorption part extraction port in the circulation system to a specific position relationship, the above problems can be solved. The present invention is completed based on such knowledge and further research.

The above-mentioned problems of the present invention are solved by the following means. [1] A stratification separation method simulating a moving layer method, which comprises a circulation system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly through piping, and the steps of separating a weakly adsorbable component relative to the adsorbent, a strongly adsorbable component, and a medium adsorbable component having an adsorption capacity between the two components contained in a stock solution by using two or more solvents; the stratification separation method simulating a moving layer method is characterized in that: a stock solution supply port F, two or more solvent supply ports D corresponding to each of the two or more solvents, an extraction port A for a weakly adsorbable portion containing the weakly adsorbable component, an extraction port B for a medium adsorbable portion containing the medium adsorbable component, and an extraction port for a strongly adsorbable portion containing the strongly adsorbable component are provided in the piping of the circulation system. The positions of the stock solution supply port F, the extraction port A, the extraction port B and the extraction port C are set as follows (a) to (c): (a) the extraction port B is set on the downstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (b) the extraction port C is set on the piping having the stock solution supply port F, or the extraction port C is set on the upstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (c) the extraction port A is set on the piping having the extraction port B, or the extraction port A is set on the downstream side of the extraction port B with at least one unit packed tower sandwiched therebetween; the stratification separation method is a simulated moving layer stratification separation method comprising sequentially repeating the following steps (A) and (B): [Step (A)] A step of supplying a stock solution from the stock solution supply port F simultaneously or separately, supplying two or more kinds of solvents from the two or more solvent supply ports D, and extracting a weakly adsorbable portion from the extraction port A, a medium adsorbable portion from the extraction port B, and a strongly adsorbable portion from the extraction port C simultaneously or separately; [Step (B)] A step of moving the stock solution supply port F, the solvent supply port D, the extraction port A, the extraction port B, and the extraction port C toward the downstream side while maintaining their relative positional relationship after the step (A) is completed. [2] A simulated moving layer analysis and separation method as described in [1], wherein the step (A) is composed of a plurality of sub-steps; the plurality of sub-steps include: a sub-step of supplying a raw liquid; and a sub-step of not supplying a raw liquid. [3] A simulated moving layer separation method as described in [1] or [2], wherein the extraction port C is arranged on the downstream side of the solvent supply port D1 for supplying the solvent d1 with the strongest desorption force among two or more solvents; at least one unit packed tower is arranged between the solvent supply port D1 and the extraction port C; and in the step (A), while the solvent d1 is supplied, the same amount of the strongly adsorbent portion as the supply amount of the solvent d1 is extracted from the extraction port C. [4] A stratification separation method using a simulated moving layer method as described in any one of [1] to [3], wherein the extraction port B is disposed on the downstream side of a solvent supply port D2 for supplying a solvent d2 having the second strongest desorption force among two or more solvents; at least one unit packed tower is disposed between the solvent supply port D2 and the extraction port B; and in the step (A), a time band is provided during the supply of the solvent d2, during which an amount of a medium adsorbent portion equal to the amount of the solvent d2 supplied is extracted from the extraction port B. [5] A stratification separation method using a simulated moving layer method as described in any one of [1] to [4], wherein 4 to 6 solvents having different desorption forces are used. [6] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 4 or more unit packed towers, and the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower, and in addition, using the two or more solvents, the following sub-steps (A1-1), (A2-1) and (A3-1) are performed in the step (A): <Sub-step (A1-1)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, supplying the stock solution from the stock solution supply port F, taking the downstream end of the segment 4 as the extraction port A, and extracting the weakly adsorbent portion from the extraction port A, thereby making the desorption force of the solvent flowing through the segment 1 the strongest, making the desorption force of the solvent flowing through the segment 2 weaker than the desorption force of the solvent flowing through the segment 1, and making the desorption force of the solvent flowing through the segments 3 and 4 weaker than the desorption force of the solvent flowing through the segment 2; ＜Sub-step (A2-1)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the upstream end of the zone 3 is used as the solvent supply port D-III, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2; ＜Sub-step (A3-1)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the downstream end of the zone 3 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream end of the zone 4 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 4 weaker than the desorption force of the solvent flowing through the zones 2 and 3. [7] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 4 or more unit packed towers, and the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and further, using the 2 or more solvents, the following sub-steps (A1-2), (A2-2) and (A3-2) are performed in the step (A): <Sub-step (A1-2)> The upstream end of the zone 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the zone 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The downstream end of the zone 4 is used as the extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, and making the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zones 1 and 2; <Sub-step (A2-2)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the zone 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the zone 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the zone 3 is used as the solvent supply port D-II. Supplying the solvent d-III from the solvent supply port D-III and extracting the weakly adsorbed portion from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2; <Sub-step (A3-2)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II in the sub-step (A2-2), the downstream side end of the zone 3 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream side end of the zone 4 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 4 weaker than the desorption force of the solvent flowing through the zones 2 and 3. [8] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 5 or more unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and in addition, the two or more solvents are used, and the following sub-steps (A1-3), (A2-3) and (A3-3) are performed in step (A): <Sub-step (A1-3)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end of the interval 5 is used as The extraction port A extracts the weakly adsorbent portion from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, making the desorption force of the solvent flowing through the zone 3 the same as or weaker than the desorption force of the solvent flowing through the zones 1 and 2, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A2-3)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the zone 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the zone 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; ＜Sub-steps (A3-3)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II in the sub-step (A2-3), the downstream side end of the interval 4 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream side end of the interval 5 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the interval 1 the strongest, the desorption force of the solvent flowing through the intervals 2, 3 and 4 weaker than the desorption force of the solvent flowing through the interval 1, and the desorption force of the solvent flowing through the interval 5 weaker than the desorption force of the solvent flowing through the intervals 2, 3 and 4. [9] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 7 or more unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and further, using the two or more solvents, the following sub-steps (A1-4), (A2-4) and (A3-4) are performed in the step (A): <Sub-step (A1-4)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end of the interval 5 is used as The extraction port A extracts the weakly adsorbent portion from the extraction port A, thereby making the desorption force of the solvent flowing through the sections 1 and 2 the strongest, making the desorption force of the solvent flowing through the section 3 the same as or weaker than the desorption force of the solvent flowing through the sections 1 and 2, and making the desorption force of the solvent flowing through the sections 4 and 5 weaker than the desorption force of the solvent flowing through the section 3; <Sub-step (A2-4)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 4 is used as the solvent supply port D-IV, supplying the solvent d-IV from the solvent supply port D-IV, and extracting the weakly adsorbent portion from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; <Sub-step (A3-4)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II in the sub-step (A2-4), the downstream side end of the interval 4 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream side end of the interval 5 is used as the solvent supply port D-V, the solvent d-V is supplied from the solvent supply port D-V, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the interval 1 the strongest, the desorption force of the solvent flowing through the intervals 2, 3 and 4 weaker than the desorption force of the solvent flowing through the interval 1, and the desorption force of the solvent flowing through the interval 5 weaker than the desorption force of the solvent flowing through the intervals 2, 3 and 4. [10] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 5 or more unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and further, using the two or more solvents, the following sub-steps (A1-5), (A2-5) and (A3-5) are performed in the step (A): <Sub-step (A1-5)> The upstream end of the zone 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F; the upstream end of the zone 4 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III; the downstream end of the zone 5 is used as the extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 3 the strongest, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A2-5)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the zone 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the zone 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; ＜Sub-steps (A3-5)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the downstream end of the zone 4 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream end of the zone 5 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4. [11] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 5 or more unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and in addition, using the two or more solvents, the following sub-steps (A1-6), (A2-6) and (A3-6) are performed in the step (A): <Sub-step (A1-6)> The upstream end of the zone 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The downstream end of the zone 3 is used as the extraction port B, and the medium adsorption part is extracted from the extraction port B. The upstream end of the zone 4 is used as the solvent supply port D-IV, and the solvent d-IV is supplied from the solvent supply port D-IV. The downstream end of the zone 5 is used as the extraction port A, and the weak adsorption part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1, 2 and 3 the strongest, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 1, 2 and 3; <Sub-step (A2-6)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the zone 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the zone 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the zone 4 is used as the solvent supply port D-II. I, supplying the solvent d-III from the solvent supply port D-III and extracting the weakly adsorbent portion from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A3-6)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the upstream end of the zone 2 is used as the solvent supply port D-II, the solvent d-II is supplied from the solvent supply port D-II, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3. [12] A simulated moving layer separation method as described in any one of [1] to [5], wherein the circulation system has 5 or more unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner such that each section has at least 1 unit packed tower, and in addition, using the two or more solvents, the following sub-steps (A1-7), (A2-7) and (A3-7) are performed in the step (A): <Sub-step (A1-7)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D-III, and the solvent is supplied from the extraction port C. The solvent d-III is supplied to the port D-III, and the downstream end of the zone 5 is used as the extraction port A. The weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; ＜Sub-step (A2-7)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the upstream end of the zone 2 is used as the solvent supply port D-II, the solvent d-II is supplied from the solvent supply port D-II, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; ＜Sub-step (A3-7)＞ The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the downstream end of the zone 4 is used as the extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream end of the zone 5 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4.〔13〕 A simulated moving layer chromatographic separation system uses a circulation system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly through piping, and separates weakly adsorbable components, strongly adsorbable components, and medium adsorbable components with adsorption between the two components contained in a stock solution by using two or more solvents; the simulated moving layer chromatographic separation system is characterized in that: in the piping of the circulation system, A stock solution supply port F, two or more solvent supply ports D corresponding to the two or more solvents, an extraction port A for a weakly adsorbable portion containing the weakly adsorbable component, an extraction port B for a medium adsorbable portion containing the medium adsorbable component, and an extraction port C for a strong adsorbable portion containing the strong adsorbable component are provided; the stock solution supply port F, the extraction port A, the extraction port B, and the extraction port C are arranged in the following positions (a) to (c): (a) The extraction port B is disposed on the downstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (b) The extraction port C is disposed on the piping having the stock solution supply port F, or the extraction port C is disposed on the upstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (c) The extraction port A is disposed on the piping having the extraction port B, or the extraction port A is disposed on the downstream side of the extraction port B with at least one unit packed tower sandwiched therebetween; The stratification separation system is a simulated moving layer stratification separation system having a mechanism for sequentially repeating the following steps (A) and (B): [Step (A)] The step of supplying the stock solution from the stock solution supply port F simultaneously or separately, supplying two or more kinds of solvents from the two or more solvent supply ports D, and extracting the weakly adsorbent part from the extraction port A, the medium adsorbent part from the extraction port B, and the strongly adsorbent part from the extraction port C simultaneously or separately; [Step (B)] After the step (A) is completed, the step of moving the stock solution supply port F, the solvent supply port D, the extraction port A, the extraction port B and the extraction port C to the downstream side while maintaining their relative positional relationship.

In this specification, the terms "upstream" and "downstream" are applicable to the flow direction of the fluid in the circulation system. That is, for a certain part of the circulation system, the so-called "upstream side" refers to the side where the fluid flows to the part, and the so-called "downstream side" refers to the side where the fluid flows out of the part. In this specification, the so-called "strongly adsorbable component" refers to a component with a stronger adsorption force relative to the adsorbent among the multiple components contained in the stock solution; the so-called "weakly adsorbable component" refers to a component with a weaker adsorption force relative to the adsorbent among the multiple components contained in the stock solution; the so-called "medium adsorbable component" refers to a component whose adsorption force relative to the adsorbent is weaker than the above-mentioned strongly adsorbable component and whose adsorption force relative to the adsorbent is stronger than the above-mentioned weakly adsorbable component. That is, the terms "strong adsorption", "medium adsorption" and "weak adsorption" show the relative strength of the adsorption force of each component contained in the stock solution when comparing the adsorption force of the adsorbent. The above-mentioned "strong adsorption component", "medium adsorption component" and "weak adsorption component" can each be composed of a single component or a plurality of components. In addition, the adsorption forces of the plurality of components can be the same or different. The grouping of "strong adsorption component", "medium adsorption component" and "weak adsorption component" of each component in the stock solution can be appropriately set according to the purpose. For example, in the case where the stock solution contains four components, the two components with the strongest adsorption force relative to the adsorbent can be assigned the position of the strong adsorption component, the component with the third strongest adsorption force relative to the adsorbent can be assigned the position of the medium adsorption component, and the component with the weakest adsorption force relative to the adsorbent can be assigned the position of the weak adsorption component. In addition, the component with the strongest adsorption force relative to the adsorbent can be assigned the position of the strong adsorption component, the components with the second strongest adsorption force relative to the adsorbent and the third strongest adsorption force relative to the adsorbent can be assigned the position of the medium adsorption component, and the component with the weakest adsorption force relative to the adsorbent can be assigned the position of the weak adsorption component. In addition, the component with the strongest adsorption force relative to the adsorbent can be assigned the status of a strongly adsorbent component, the component with the second strongest adsorption force relative to the adsorbent can be assigned the status of a medium adsorbent component, and the component with the third strongest adsorption force relative to the adsorbent and the weakest component can be assigned the status of a weakly adsorbent component. The original solution containing more than 5 components can also be separated and purified based on various components. In the present invention, the so-called "desorption force" of the elution solution refers to the strength of the action that instructs the component adsorbed by the adsorbent to desorb from the adsorbent. [Effects of the invention]

According to the simulated shifting layer analysis separation method of the present invention, the amount of adsorbent used can be suppressed while the purification target component in the stock solution can be extracted with high purity. In addition, the simulated shifting layer analysis separation system of the present invention can be appropriately used to implement the simulated shifting layer analysis separation method of the present invention.

The preferred implementation of the simulated moving layer analysis and separation method of the present invention (hereinafter also referred to as "the method of the present invention") is described below.

The method of the present invention is implemented by a circulation system in which a plurality of unit packed towers filled with adsorbent are connected in series and endlessly through piping. The circulation system body used for simulating the moving bed method is well known to the public. For example, reference can be made to Japanese Patent Publication No. 2009-36536 or Japanese Patent No. 4606092. The circulation system is described below with diagrams, but the present invention is not limited to such implementation modes except for the contents defined by the present invention. In addition, the diagrams disclosed below are explanatory diagrams for making the present invention easier to understand. The dimensions or relative size relationships of each structure are sometimes changed in size for the convenience of explanation, and the actual relationship is not displayed in its original state. In addition, except for the matters defined by the present invention, the shapes, relative positional relationships, etc. disclosed in the drawings are not limited. In addition, conditions other than those defined by the present invention, such as the capacity of a unit packed column, the cross-sectional area or length of the pipe, the flow rate of the liquid supplied to the circulation system, etc., can be appropriately set according to the purpose.

A preferred embodiment of the circulation system used in the method of the present invention is disclosed in FIG1 . The circulation system 100 shown in FIG1 has four unit-packed towers (columns) (unit-packed towers 10a, 10b, 10c, and 10d) filled with adsorbent Ab. The outlet of each unit-packed tower is connected to the inlet of the adjacent unit-packed tower through a pipe 111, and each unit-packed tower is connected in series to form a whole. Then, the outlet of the last unit-packed tower (e.g., unit-packed tower 10d) is connected to the inlet of the first unit-packed tower (e.g., unit-packed tower 10a) through a pipe 111, and all the unit-packed towers are connected in an endless shape (ring shape). With such a structure, the fluid can be circulated in the circulation system 100. The unit packed towers 10a to 10d may be identical or different in shape, size, and amount of adsorbent filled therein. The unit packed towers 10a to 10d may be identical (more preferably identical) in shape, size, and amount of adsorbent filled therein.

A circulation pump P1 for circulating the fluid in the direction of the arrow may be provided in the circulation system 100. The circulation pump P1 is preferably a metering pump. In addition, in the circulation system 100, a pipe 111 between two adjacent unit packed towers is provided with stop valves R1, R2, R3, and R4 that can block the flow of the fluid to the unit packed tower on the downstream side.

Between each shutoff valve R1 to R4 and the outlet of each unit packed tower 10a to 10d located on the upstream side thereof, weakly adsorbable part extraction pipelines 2a, 2b, 2c, and 2d are respectively arranged in a branching manner to extract the part containing more weakly adsorbable components relative to the adsorbent Ab (in this specification, "weakly adsorbable part relative to the adsorbent Ab" is also referred to as "weakly adsorbable part" for short). Weakly adsorbable part extraction valves A1, A2, A3, and A4 that can open and close each weakly adsorbable part extraction pipeline are respectively arranged on each weakly adsorbable part extraction pipeline 2a, 2b, 2c, and 2d. Each weakly adsorbable part extraction pipeline 2a, 2b, 2c, and 2d merges and converges into a weakly adsorbable part converging pipe 2J.

Similarly, between each of the block valves R1 to R4 and the outlet of each of the unit packed towers 10a to 10d located on the upstream side thereof, there are provided in a branched manner intermediate adsorbent part extraction pipelines 3a, 3b, 3c, and 3d for extracting a portion containing a relatively large amount of intermediate adsorbent components relative to the adsorbent Ab (in this specification, "intermediate adsorbent portion relative to the adsorbent Ab" is also referred to as "intermediate adsorbent portion" for short). In each of the intermediate adsorbent part extraction pipelines 3a, 3b, 3c, and 3d, intermediate adsorbent part extraction valves B1, B2, B3, and B4 that can open and close each intermediate adsorbent part extraction pipeline are provided. Each of the intermediate adsorbent part extraction pipelines 3a, 3b, 3c, and 3d merges and is collected in a intermediate adsorbent part converging pipe 3J.

Similarly, between each of the block valves R1 to R4 and the outlet of each of the unit packed towers 10a to 10d located on the upstream side thereof, strongly adsorbed portion extraction pipelines 4a, 4b, 4c, and 4d are provided in a branched manner to extract a portion containing a relatively large amount of strongly adsorbed components relative to the adsorbent Ab (in this specification, "strongly adsorbed portion relative to the adsorbent Ab" is also referred to as "strongly adsorbed portion" for short). Strongly adsorbed portion extraction valves C1, C2, C3, and C4 that can open and close each strongly adsorbed portion extraction pipeline are provided in each of the strongly adsorbed portion extraction pipelines 4a, 4b, 4c, and 4d. Each of the strongly adsorbed portion extraction pipelines 4a, 4b, 4c, and 4d merges and converges into a strongly adsorbed portion converging pipe 4J.

In the step (A) described later, any one of the weak adsorption part extraction valves A1, A2, A3, and A4 is in an open valve state. The connection portion between the weak adsorption part extraction line of the weak adsorption part extraction valve that is set to be open and the pipe 111 is the weak adsorption part extraction port A in the step (A) described later. In addition, in the step (A) described later, any one of the medium adsorption part extraction valves B1, B2, B3, and B4 is in an open valve state. The connection portion between the medium adsorption part extraction line of the medium adsorption part extraction valve that is set to be open and the pipe 111 is the medium adsorption part extraction port B in the step (A) described later. In addition, in step (A) described later, any one of the strong adsorption part extraction valves C1, C2, C3, and C4 is in an open state. The connection portion between the strong adsorption part extraction pipeline provided with the open strong adsorption part extraction valve and the pipe 111 is the strong adsorption part extraction port C in step (A) described later.

In the circulation system 100, a safety valve (or pressure reducing valve) not shown in the figure may be installed at an appropriate position to prevent excessive pressure rise in the circulation system 100. In addition, check valves T1, T2, T3, and T4 for preventing backflow should be installed between two adjacent unit packed towers.

In the circulation system 100, as shown in FIG1, a structure is constructed to supply a raw liquid 117 stored in a raw liquid tank 116. In addition, in the circulation system 100, a structure is constructed to supply two or more kinds of solvents. In FIG1, as an example, a state of supplying four kinds of solvents is shown. The raw liquid 117 is supplied through the raw liquid supply pipeline 11 by using a raw liquid supply pump P2 that can control the supply flow rate. The raw liquid supply pump P2 is preferably a metering pump. The raw liquid supply pipeline 11, as shown in FIG1, has a structure of "branching out four raw liquid supply branch pipelines 11a, 11b, 11c, and 11d, and the raw liquid can be supplied to the inlet of each unit filling tower 10a, 10b, 10c, and 10d through each raw liquid supply branch pipeline 11a, 11b, 11c, and 11d." In each of the raw liquid supply branch pipelines 11a, 11b, 11c, and 11d, openable and closable raw liquid supply valves F1, F2, F3, and F4 are provided, and the raw liquid is supplied to the unit packed tower connected downstream thereof through the raw liquid supply branch pipeline having the openable raw liquid supply valve. In the step (A) described later, any one of the raw liquid supply valves F1, F2, F3, and F4 is in the open valve state. The connection portion between the raw liquid supply branch pipeline having the open valve raw liquid supply valve and the piping 111 is the raw liquid supply port F in the step (A) described later.

FIG1 shows the state of supplying four kinds of solvents with different desorption forces. The solvent 9a stored in the solvent tank 8a is supplied to the solvent supply pipeline 12 by the solvent supply pump P3 with controllable supply flow rate. The solvent 9b stored in the solvent tank 8b is supplied to the solvent supply pipeline 13 by the solvent supply pump P4 with controllable supply flow rate. The solvent 9c stored in the solvent tank 8c is supplied to the solvent supply pipeline 14 by the solvent supply pump P5 with controllable supply flow rate. Furthermore, the solvent 9d stored in the solvent tank 8d is supplied to the solvent supply pipeline 15 by the solvent supply pump P6 with controllable supply flow rate. The solvent supply pumps P3 to P6 are preferably quantitative pumps. As shown in FIG. 1 , the solvent supply pipeline 12 is structured such that “four solvent supply branch pipelines 12a, 12b, 12c, and 12d are branched, and the solvent can be supplied to the inlet of each unit packed tower 10a, 10b, 10c, and 10d through each solvent supply branch pipeline 12a, 12b, 12c, and 12d”. On each solvent supply branch pipeline 12a, 12b, 12c, and 12d, an openable and closable solvent supply valve E1a, E2a, E3a, and E4a are provided, and the solvent is supplied to the unit packed tower connected downstream thereof through the solvent supply branch pipeline having the openable and closable solvent supply valve. Similarly, the solvent supply pipeline 13 branches out into four solvent supply branch pipelines 13a, 13b, 13c, and 13d, the solvent supply pipeline 14 branches out into four solvent supply branch pipelines 14a, 14b, 14c, and 14d, and the solvent supply pipeline 15 branches out into four solvent supply branch pipelines 15a, 15b, 15c, and 15d, thereby forming a structure that can supply each solvent to the inlet of each unit packed tower 10a, 10b, 10c, and 10d. The solvent supply branch lines 13a, 13b, 13c, and 13d are provided with openable and closable solvent supply valves E1b, E2b, E3b, and E4b, respectively. The solvent supply branch lines 14a, 14b, 14c, and 14d are provided with openable and closable solvent supply valves E1c, E2c, E3c, and E4c, respectively. The solvent supply branch lines 15a, 15b, 15c, and 15d are provided with openable and closable solvent supply valves E1d, E2d, E3d, and E4d, respectively. In the step (A) described later, the connection portion between the solvent supply branch line provided with the openable solvent supply valve and the pipe 111 is the solvent supply port D. In the method of the present invention, since two or more solvents are used, in the step (A) described later, the solvent supply valve to be opened is plural. Therefore, in the step (A) described later, the number of solvent supply ports D will correspond to the number of types of solvents used (two or more).

Next, the operation of the circulation system when the method of the present invention is implemented using the above-mentioned circulation system is explained, but the present invention is not limited to such implementation forms except for the contents limited by the present invention. In the method of the present invention, the circulation system makes the positions of the stock solution supply port F, the weak adsorption part extraction port A, the medium adsorption part extraction port B, and the strong adsorption part extraction port C form the following relationship (a) to (c). That is, in the repetition of the steps (A) and (B) described later, the relative position relationship between the stock solution supply port F, the weak adsorption part extraction port A, the medium adsorption part extraction port B, and the strong adsorption part extraction port C always satisfies the above (a) to (c).

(a) The middle adsorptive fraction extraction port B is provided on the downstream side of the stock solution supply port F with at least one unit packed tower interposed therebetween.

(b) The strong adsorption part extraction port C is set in the piping having the stock solution supply port F, or the strong adsorption part extraction port C is set on the upstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween. Here, "the strong adsorption part extraction port C is set in the piping having the stock solution supply port F" means that no unit packed tower is arranged between the strong adsorption part extraction port C and the stock solution supply port F. In addition, when "the strong adsorption part extraction port C is set in the piping having the stock solution supply port F", the strong adsorption part extraction port C is set on the upstream side of the same piping as the stock solution supply port F. This conforms to the overall relationship between the extraction port and the supply port set in the same piping. That is, in a circulation system, when a certain extraction port and a supply port are set in the same pipe (when the extraction port and the supply port are set in a manner that does not sandwich a unit packed tower), the extraction port is arranged on the upstream side of the same pipe compared to the supply port. This is to prevent the liquid supplied from being extracted from the extraction port before it reaches the unit packed tower downstream. The above (b) is preferably in the form of "arranging the strong adsorption part extraction port C on the upstream side of the stock liquid supply port F sandwiching at least one unit packed tower".

(c) The weak adsorption part extraction port A is provided in the piping having the medium adsorption part extraction port B, or the weak adsorption part extraction port A is provided on the downstream side of the medium adsorption part extraction port B with at least one unit packed tower sandwiched therebetween. Here, "the weak adsorption part extraction port A is provided in the piping having the medium adsorption part extraction port B" means that no unit packed tower is provided between the weak adsorption part extraction port A and the medium adsorption part extraction port B. The above (c) is preferably in the form of "the weak adsorption part extraction port A is provided on the downstream side of the medium adsorption part extraction port B with at least one unit packed tower sandwiched therebetween".

In the method of the present invention, the above-mentioned circulation system is used to repeat the following steps (A) and (B) in sequence.

[Step (A)] A step of supplying a stock solution from a stock solution supply port F at the same time or separately, supplying two or more solvents from two or more solvent supply ports D, and extracting a weakly adsorbed portion from a weakly adsorbed portion extraction port A at the same time or separately, extracting a medium adsorbed portion from a medium adsorbed portion extraction port B, and extracting a strongly adsorbed portion from a strongly adsorbed portion extraction port C at the same time or separately. Here, in step (A), the sum of the amount of the stock solution supplied from the stock solution supply port F and the amount of the solvent supplied from the solvent supply port D is equal to the sum of the amount of the weakly adsorbed portion extracted from the weakly adsorbed portion extraction port A, the amount of the medium adsorbed portion extracted from the medium adsorbed portion extraction port B, and the amount of the strongly adsorbed portion C extracted from the strongly adsorbed portion extraction port C. That is, when liquid is supplied to the circulation system, the same amount of liquid is extracted from the circulation system. To be more specific, when liquid is supplied from a certain supply port (X) and extracted from a certain extraction port (Y) on the downstream side thereof, if liquid is not supplied from a place further downstream than X and further upstream than Y, the amount of liquid extracted from Y is the same as the amount of liquid supplied from X. On the other hand, if liquid is supplied from a place further downstream than X and further upstream than Y, the amount of liquid extracted from Y is the same as the sum of the amount of liquid supplied from X and the amount of liquid supplied from a place further downstream than X and further upstream than Y. For example, in the sub-step (A1-1) of FIG. 2 , the amount of the solvent supplied from the solvent supply port D1 is the same as the amount of the strongly adsorbable part extracted from the strongly adsorbable part extraction port C. In addition, in the same sub-step (A1-1), the total amount of the solvent supplied from the solvent supply port D2 and the amount of the stock solution supplied from the stock solution supply port F is the same as the amount of the weakly adsorbable part extracted from the weakly adsorbable part extraction port A. In addition, the above-mentioned "simultaneous or separate supply" means supplying without setting a time difference (without staggering the supply timing) or supplying with setting a time difference (with staggering the supply timing). Among them, in step (A), when two or more supply ports for supplying two or more liquids are arranged in the same pipe (when the two or more supply ports are arranged in the pipe without sandwiching a unit packed tower, and different liquids are supplied from each of the two or more supply ports), the supply of the two or more liquids will not be implemented at the same time. That is, in step (A), the supply of the two or more liquids is implemented in different sub-steps. Similarly, in step (A), when two or more extraction ports for extracting two or more parts are arranged in the same pipe (when the two or more extraction ports are arranged in the pipe without sandwiching a unit packed tower, and different parts are extracted from each of the two or more extraction ports), the extraction of the two or more parts will not be implemented at the same time. That is, in step (A), the extraction of the two or more parts is performed in different sub-steps.

[Step (B)] After the step (A) is completed, the raw liquid supply port F, the elution liquid supply port D, the weak adsorption part extraction port A, the medium adsorption part extraction port B, and the strong adsorption part extraction port C are moved downstream while maintaining their relative positional relationship. The movement to the downstream side means that the raw liquid supply port F, the elution liquid supply port D, the weak adsorption part extraction port A, the medium adsorption part extraction port B, and the strong adsorption part extraction port C are instructed to move downstream by 1 unit of the unit filling tower while maintaining their relative positional relationship. For example, when in step (A), the connection part between the stock solution supply branch line provided with the stock solution supply valve F1 and the piping 111 is the stock solution supply port F, by step (B), the stock solution supply port F is moved to the connection part between the stock solution supply branch line provided with the stock solution supply valve F2 and the piping 111. This is also the case with the elution supply port D, the weakly adsorbent part extraction port A, the medium adsorbent part extraction port B, and the strong adsorbent part extraction port C. In addition, the movement of the above-mentioned supply ports and extraction ports to the downstream side of the unit packed tower by one unit can be implemented by adjusting the opening and closing of various pumps or various valves arranged in the circulation system. By carrying out step (B), in the following step (A) [referred to as step (A2)], the supply and extraction of liquid are carried out for each unit packed tower one unit downstream of step (A1) in the same manner as carried out for each unit packed tower in step (A) [referred to as step (A1)] before step (B).

The above step (A) is preferably composed of a plurality of sub-steps. At this time, the supply of the stock solution from the stock solution supply port F, the supply of two or more kinds of elution solutions from the two or more elution solution supply ports D, the extraction of the weakly adsorbable part from the weakly adsorbable part extraction port A, the extraction of the medium adsorbable part from the medium adsorbable part extraction port B, and the extraction of the strong adsorbable part from the strong adsorbable part extraction port C, which sub-step is implemented can be appropriately set according to the purpose within the scope that does not impair the efficacy of the present invention. In particular, in the method of the present invention, the above step (A) is preferably composed of a sub-step of supplying the stock solution and a sub-step of not supplying the stock solution. That is, in step (A), there is preferably a time when the stock solution is supplied and a time when the stock solution is not supplied.

In step (A), two or more solvents are supplied. The supply port of the solvent with the strongest desorption force among the two or more solvents is preferably set in the pipe on the upstream side of the unit packed tower relative to the strong adsorption part extraction port C. The supply ports of the remaining solvents and even the stock solution supply port are preferably set in the same pipe as the supply port of the solvent with the strongest desorption force, or, compared with the supply port of the solvent with the strongest desorption force, are set on the upstream side of the unit packed tower. The phrase "being provided in the same pipe as the supply port of the solvent with the strongest desorption force, or being provided on the upstream side of the unit packed tower compared to the supply port of the solvent with the strongest desorption force" means that a supply port for a solvent other than the strongest solvent or even a supply port for the stock solution is provided in any of the pipes from the pipe having the supply port of the strongest solvent to the upstream side until the pipe having the extraction port C of the strongly adsorbent part is provided (in other words, the pipe having the extraction port C of the strongly adsorbent part is provided) The piping of the extraction port C goes downstream from the starting point until it reaches the piping where the supply port of the strongest solvent is set. In any of the pipings in between, a supply port for solvents other than the strongest solvent or even a supply port for the stock solution is set. In other words, when two or more unit packed towers are set between the supply port of the strongest solvent and the extraction port C of the strong adsorption part, a supply port for solvents or even a supply port for the stock solution is not set in the piping connecting the two or more unit packed towers. At this time, if there are two or more solvents other than the strongest solvent, part of the two or more supply ports corresponding to the solvents may be set in the same piping, or each may be set in a separate piping sandwiching the unit packed towers. In addition, the supply port for solvents other than the strongest solvent and the stock solution supply port may also be set in the same piping. In addition, it is a preferred embodiment of the present invention that one of the supply ports for the solvents other than the solvent with the strongest desorption force is provided in the same pipe as the supply port for the solvent with the strongest desorption force.

In step (A), it is preferable that "while supplying the solvent with the strongest desorption force among two or more solvents, the same amount of the strongly adsorbent fraction as the supply amount of the solvent with the strongest desorption force is extracted from its downstream". In this case, it is preferable that "at least one unit packed tower is arranged between the supply port of the solvent with the strongest desorption force and the extraction port of the strongly adsorbent fraction downstream thereof, and no other supply port exists in between".

In step (A), it is preferable to set a time zone (sub-step) in which the same amount of the medium adsorbent as the supply amount of the second strongest desorbent among two or more solvents is extracted from the downstream thereof. At this time, at least one unit packed tower (preferably a plurality of unit packed towers) is arranged between the supply port of the second strongest desorbent to the outlet for extracting the medium adsorbent downstream thereof. In addition, even if there are other supply ports between the piping provided with the supply port of the second strongest desorbent to the piping provided with the outlet for extracting the medium adsorbent on the downstream side thereof, liquid is not supplied from the other supply ports during the time zone for extracting the medium adsorbent.

During the implementation of step (A), it is preferred to frequently extract the weakly adsorbable fraction. Therefore, when step (A) is composed of a plurality of sub-steps, it is also preferred to frequently extract the weakly adsorbable fraction in the plurality of sub-steps.

In the method of the present invention, it is preferred to use three or more solvents with different desorption forces, more preferably four or more solvents with different desorption forces, more preferably four to six solvents with different desorption forces, and particularly preferably four or five solvents with different desorption forces. The type of solvent is not particularly limited and is appropriately set according to its relationship with the type of adsorbent or the type of components in the stock solution. For example, when an ion exchange resin is used as an adsorbent, a plurality of solvents with different desorption forces can be prepared by changing the salt concentration of the solvent. For example, when a cation exchange resin is used, a plurality of solvents with changed NaCl concentrations can be used as two or more solvents.

The preferred implementation of the combination of sub-steps in the above step (A) is described below. The implementation of such aspects can be implemented using the system of Figure 1, or a system that can implement the objective aspects based on the system of Figure 1. In addition, the following implementation is an example of the present invention. For example, from the perspective of relative desorption force, it is also possible to prepare two or more solvents that are given the status of the solvent d-1 described below, and change the type of solvent d-1 used between the sub-steps of supplying the solvent d-1. In this regard, the same is true for solvents d-II to d-V. That is, in the present invention, when "solvent d-1" is referred to in a certain embodiment, if "solvent d-1" is used in different sub-steps in the embodiment, the "solvent d-1" used in the different sub-steps may be the same as or different from each other. This also applies to solvents d-II to d-V.

-Implementation Sample 1- In Implementation Sample 1, a circulation system having 4 or more unit packed towers is used. Then, it is assumed that "the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this Implementation Sample 1, the following sub-steps (A1-1), (A2-1) and (A3-1) are implemented in step (A). In the present invention, the phrase "implementing sub-steps X, Y, and Z in step (A)" means that step (A) includes sub-steps X, Y, and Z. The order of implementing sub-steps X, Y, and Z can be appropriately set within the scope that does not impair the effectiveness of the present invention. In addition, step (A) may also include other sub-steps besides sub-steps X, Y, and Z. Regarding the aspect of "implementing sub-steps X, Y, and Z in step (A)", typically, an aspect of implementing sub-steps X, Y, and Z in sequence as step (A) can be listed, but the present invention is not limited to this aspect. That is, the so-called "implementing sub-steps X, Y, and Z in step (A)" is not limited to the aspect of "implementing sub-step Y after sub-step X, implementing sub-step Z after sub-step Y, implementing step (B) after sub-step Z, and implementing sub-step X after step (B)". For example, in the above-mentioned aspect including other sub-steps, other sub-steps (sub-steps other than sub-steps X, Y, and Z) may be combined and inserted in at least one of the following places: before sub-step X [between step (B) and sub-step X], between sub-steps X and Y, between sub-steps Y and Z, and between sub-steps Z and step (B), within the scope that does not impair the efficacy of the present invention. Sometimes, by adjusting the supply flow rate, flow velocity, solvent strength, etc., the desired effect can be achieved even if additional sub-steps other than sub-steps X, Y, and Z are inserted. This will be easily understood by practitioners in the field who have access to this specification.

<Sub-step (A1-1)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the solvent supply port D-II. The stock solution supply port F is used to supply the stock solution, and the downstream end of the zone 4 is used as the weakly adsorbent part extraction port A, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2. In the sub-step (A1-1), the supply of each liquid and the extraction of each part are continuously implemented (that is, in the sub-step (A1-1), the supply of each liquid and the extraction of each part are all implemented continuously). In addition, in the present invention and even in this specification, the strength of the desorption of the solvent described in each sub-step refers to the strength of the desorption of the solvent in the sub-step, and does not refer to the strength of the desorption of the solvent in the different sub-steps. For example, when it is stated that in a sub-step constituting step (A), the desorption force of the solvent flowing through zone 1 is the strongest, and it is stated that in another sub-step constituting step (A), the desorption force of the solvent flowing through zone 1 is also the strongest, the desorption force of the solvent flowing through zone 1 in the one sub-step and the desorption force of the solvent flowing through zone 1 in the other sub-step may be the same or different.

<Sub-step (A2-1)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the upstream end of the segment 3 is used as the solvent supply port D-III, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through segment 1 the strongest, making the desorption force of the solvent flowing through segment 2 weaker than the desorption force of the solvent flowing through segment 1, and making the desorption force of the solvent flowing through segments 3 and 4 weaker than the desorption force of the solvent flowing through segment 2. The solvent supply port D-III in the sub-step (A2-1) is provided in the same pipe as the stock solution supply port F in the above-mentioned sub-step (A1-1).

<Sub-step (A3-1)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II, the downstream end of the segment 3 is used as the medium adsorbent portion extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream end of the segment 4 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through segment 1 the strongest, making the desorption force of the solvent flowing through segments 2 and 3 weaker than the desorption force of the solvent flowing through segment 1, and making the desorption force of the solvent flowing through segment 4 weaker than the desorption force of the solvent flowing through segments 2 and 3.

The desorption force of the solvent flowing through the sections 3 and 4 in the above sub-step (A2-1) is preferably the same as the desorption force of the solvent flowing through the section 4 in the above sub-step (A3-1).

As an example of the above sub-step (A1-1), a sub-step for implementing the following sub-step (A1-1ex) can be listed, but the above sub-step (A1-1) is not limited to the sub-step (A1-1ex). <Sub-step (A1-1ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The downstream end of the interval 4 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above sub-step (A2-1), a sub-step of implementing the following sub-step (A2-1ex) can be listed, but the above sub-step (A2-1) is not limited to sub-step (A2-1ex). ＜Sub-step (A2-1ex)＞ The solvent d1 is supplied from the solvent supply port D1, the strongly adsorbent portion is extracted from the strongly adsorbent portion extraction port C, the solvent d2 is supplied from the solvent supply port D2, the upstream end of the interval 3 is used as the solvent supply port D3, the solvent d3 with the weakest desorption force among the four solvents is supplied from the solvent supply port D3, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A. The solvent supply port D3 in the sub-step (A2-1ex) is provided in the same pipe as the stock solution supply port F in the above-mentioned sub-step (A1-1).

As an example of the above-mentioned sub-step (A3-1), a sub-step of implementing the following sub-step (A3-1ex) can be listed, but the above-mentioned sub-step (A3-1) is not limited to sub-step (A3-1ex). ＜Sub-step (A3-1ex)＞ The solvent d1 is supplied from the solvent supply port D1, the strongly adsorbed part is extracted from the strongly adsorbed part extraction port C, the solvent d2 is supplied from the solvent supply port D2, the downstream end of the interval 3 is used as the medium adsorbed part extraction port B, the medium adsorbed part is extracted from the extraction port B, the upstream end of the interval 4 is used as the solvent supply port D4, the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4, and the weakly adsorbed part is extracted from the weakly adsorbed part extraction port A.

Taking the embodiment in which each compartment has a unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-1ex), (A2-1ex) and (A3-1ex) is shown in FIG2. In FIG2, a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower (numbered sequentially from the left). After the step (A) of sequentially performing the above sub-steps (A1-1ex), (A2-1ex) and (A3-1ex) is completed, the stock solution supply port F, the elution supply port D, the weakly adsorbent part extraction port A, the medium adsorbent part extraction port B and the strong adsorbent part extraction port C are moved downstream while maintaining their relative positional relationship through step (B). Then, a flow chart of the state of sequentially performing the above sub-steps (A1-1ex), (A2-1ex) and (A3-1ex) is shown in FIG3. The unit packed towers arranged in each compartment shown in FIG2 are displaced one by one toward the downstream side in FIG3. At this point, consider "starting from step (A) shown in Figure 2, and then performing step (B)" as one set of steps. By performing four sets of steps, we will return to the state shown in Figure 2.

-Implementation Example 2- Implementation Example 2 is also the same as Implementation Example 1, using a circulation system having more than 4 unit packed towers. Then, it is assumed that "the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this Implementation Example 2, the following sub-steps (A1-2), (A2-2) and (A3-2) are sequentially implemented as step (A).

<Sub-step (A1-2)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The downstream end of the interval 4 is used as the weakly adsorbable part extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the intervals 1 and 2 the strongest, and making the desorption force of the solvent flowing through the intervals 3 and 4 weaker than the desorption force of the solvent flowing through the intervals 1 and 2.

<Sub-step (A2-2)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the solvent supply port D-II. The end is a solvent supply port D-III, and a solvent d-III is supplied from the solvent supply port D-III, and a weakly adsorbable portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2. The solvent supply port D-I in the sub-step (A2-2) is provided in the same pipe as the solvent supply port D-II in the above sub-step (A1-2). In addition, the solvent supply port D-III is provided in the same pipe as the stock solution supply port F. The desorption force of the solvent flowing through the interval 1 in the sub-step (A2-2) is preferably stronger than the desorption force of the solvent flowing through the interval 1 in the above sub-step (A1-2).

<Sub-step (A3-2)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II in sub-step (A2-2), the downstream end of the interval 3 is used as the medium adsorbent portion extraction port B, the medium adsorbent portion is extracted from the extraction port B, and the upstream end of the interval 4 is used as the solvent supply port The solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 4 weaker than the desorption force of the solvent flowing through the zones 2 and 3.

As an example of the above-mentioned sub-step (A1-2), a sub-step of implementing the following sub-step (A1-2ex) can be listed, but the above-mentioned sub-step (A1-2) is not limited to the sub-step (A1-2ex). ＜Sub-step (A1-2ex)＞ The upstream end of the interval 1 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2, the upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F, and the downstream end of the interval 4 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above sub-step (A2-2), a sub-step for implementing the following sub-step (A2-2ex) can be listed, but the above sub-step (A2-2) is not limited to the sub-step (A2-2ex). <Sub-step (A2-2ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The upstream end of the interval 3 is used as the solvent supply port D3, and the solvent d3 with the weakest desorption force among the four solvents is supplied from the solvent supply port D3, and the weak adsorption part is extracted from the weak adsorption part extraction port A. The solvent supply port D1 in the sub-step (A2-2ex) is provided in the same pipe as the solvent supply port D2 in the above sub-step (A1-2ex). In addition, the solvent supply port D3 is provided in the same pipe as the stock solution supply port F in the above sub-step (A1-2ex).

As an example of the above sub-step (A3-2), a sub-step for implementing the following sub-step (A3-2ex) can be listed, but the above sub-step (A3-2) is not limited to the sub-step (A3-2ex). <Sub-step (A3-2ex)> The solvent d1 is supplied from the solvent supply port D1, and the strongly adsorbent portion is extracted from the strongly adsorbent portion extraction port C. The solvent d2 is supplied from the solvent supply port D2 in the sub-step (A2-2ex), and the downstream end of the interval 3 is used as the medium adsorbent portion extraction port B, and the medium adsorbent portion is extracted from the extraction port B. The upstream end of the interval 4 is used as the solvent supply port D4, and the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the embodiment in which each zone has a unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-2ex), (A2-2ex) and (A3-2ex) is shown in FIG4. In FIG4, a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower. After the step (A) of sequentially performing the above sub-steps (A1-2ex), (A2-2ex) and (A3-2ex) is completed, the stock solution supply port F, the elution supply port D, the weak adsorption part extraction port A, the medium adsorption part extraction port B and the strong adsorption part extraction port C are moved to the downstream side while maintaining their relative positional relationship through step (B), and then, the flow chart of the state of sequentially performing the above sub-steps (A1-2ex), (A2-2ex) and (A3-2ex) is shown in FIG5. The unit packed towers arranged in each compartment shown in FIG4 are shifted to the downstream side one by one in FIG5. At this time, "starting from step (A) shown in FIG4, and then performing step (B)" is regarded as one set of steps, and by performing four sets of steps, the state shown in FIG4 is returned again.

-Implementation Sample 3- Implementation Sample 3 uses a circulation system having more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this implementation sample 3, the following sub-steps (A1-3), (A2-3) and (A3-3) are sequentially implemented as step (A).

<Sub-step (A1-3)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end of the interval 5 is used as the solvent supply port D-III. The end serves as an extraction port A for the weakly adsorbent part, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the sections 1 and 2 the strongest, making the desorption force of the solvent flowing through the section 3 the same as or weaker than the desorption force of the solvent flowing through the sections 1 and 2, and making the desorption force of the solvent flowing through the sections 4 and 5 weaker than the desorption force of the solvent flowing through the section 3.

<Sub-step (A2-3)> Use the upstream end of the interval 1 as the solvent supply port D-I, and supply the solvent d-I from the solvent supply port D-I; use the downstream end of the interval 1 as the strong adsorption part extraction port C, and extract the strong adsorption part from the extraction port C; use the upstream end of the interval 2 as the solvent supply port D-II, and supply the solvent d-II from the solvent supply port D-II , the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3. The desorption force of the solvent flowing through the zone 1 in the sub-step (A2-3) is preferably stronger than the desorption force of the solvent flowing through the zone 1 in the above sub-step (A1-3). The solvent supply port D-I in the sub-step (A2-3) is provided in the same pipe as the solvent supply port D-II in the above-mentioned sub-step (A1-3).

<Sub-step (A3-3)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d-II is supplied from the solvent supply port D-II in the sub-step (A2-3), the downstream end of the interval 4 is used as the medium adsorbent portion extraction port B, the medium adsorbent portion is extracted from the extraction port B, and the upstream end of the interval 5 is used as the solvent supply port D-IV, the solvent d-IV is supplied from the solvent supply port D-IV, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4.

As an example of the above sub-step (A1-3), a sub-step for implementing the following sub-step (A1-3ex) can be listed, but the above sub-step (A1-3) is not limited to the sub-step (A1-3ex). <Sub-step (A1-3ex)> The upstream end of the interval 1 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D3, and the solvent d3 with the weakest desorption force among the four solvents is supplied from the solvent supply port D3. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above sub-step (A2-3), a sub-step for implementing the following sub-step (A2-3ex) can be listed, but the above sub-step (A2-3) is not limited to the sub-step (A2-3ex). <Sub-step (A2-3ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The solvent d3 is supplied from the solvent supply port D3, and the weak adsorption part is extracted from the weak adsorption part extraction port A. The solvent supply port D1 in the sub-step (A2-3ex) is provided in the same pipe as the solvent supply port D2 in the above sub-step (A1-3ex).

As an example of the above sub-step (A3-3), a sub-step for implementing the following sub-step (A3-3ex) can be listed, but the above sub-step (A3-3) is not limited to the sub-step (A3-3ex). <Sub-step (A3-3ex)> The solvent d1 is supplied from the solvent supply port D1, and the strongly adsorbent portion is extracted from the strongly adsorbent portion extraction port C. The solvent d2 is supplied from the solvent supply port D2 in the sub-step (A2-3ex), and the downstream end of the interval 4 is used as the medium adsorbent portion extraction port B, and the medium adsorbent portion is extracted from the extraction port B. The upstream end of the interval 5 is used as the solvent supply port D4, and the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the embodiment in which each zone has a unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-3ex), (A2-3ex) and (A3-3ex) is shown in FIG6 . In FIG6 , a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower. After the step (A) of sequentially performing the above sub-steps (A1-3ex), (A2-3ex) and (A3-3ex) is completed, the stock solution supply port F, the elution supply port D, the weak adsorption part extraction port A, the medium adsorption part extraction port B and the strong adsorption part extraction port C are moved to the downstream side while maintaining their relative positional relationship through step (B), and then, the flow chart of the state of sequentially performing the above sub-steps (A1-3ex), (A2-3ex) and (A3-3ex) is shown in FIG7. The unit packed towers arranged in each compartment shown in FIG6 are shifted to the downstream side one by one in FIG7. At this time, "starting from step (A) shown in FIG6, and then performing step (B)" is regarded as one set of steps, and by performing five sets of them, the state shown in FIG6 is returned again.

-Implementation Sample 4- Implementation Sample 4 uses a circulation system having more than 7 unit packed towers. Then, it is assumed that "the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 5 types of eluents d-I to d-V with different desorption forces are used as eluents. In this implementation sample 4, the following sub-steps (A1-4), (A2-4) and (A3-4) are sequentially implemented as step (A).

<Sub-step (A1-4)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end of the interval 5 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end serves as the extraction port A, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, making the desorption force of the solvent flowing through the zone 3 the same as or weaker than the desorption force of the solvent flowing through the zones 1 and 2, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3.

<Sub-step (A2-4)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream end of the interval 4 is used as the solvent supply port D-II. The end is used as a solvent supply port D-IV, a solvent d-IV is supplied from the solvent supply port D-IV, and a weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3. The desorption force of the solvent flowing through the zone 1 in the sub-step (A2-4) is preferably stronger than the desorption force of the solvent flowing through the zone 1 in the above sub-step (A1-4). The solvent supply port D-I in the sub-step (A2-4) is provided in the same pipe as the solvent supply port D-II in the above-mentioned sub-step (A1-4).

<Sub-step (A3-4)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the solvent d2 is supplied from the solvent supply port D2 in the sub-step (A2-4), the downstream end of the interval 4 is used as the medium adsorbent portion extraction port B, the medium adsorbent portion is extracted from the extraction port B, and the upstream end of the interval 5 is used as the solvent supply port D -V, the solvent d-V is supplied from the solvent supply port D-V, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4.

As an example of the above sub-step (A1-4), a sub-step for implementing the following sub-step (A1-4ex) can be listed, but the above sub-step (A1-4) is not limited to the sub-step (A1-4ex). <Sub-step (A1-4ex)> The upstream end of the interval 1 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the five solvents is supplied from the solvent supply port D2. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D3, and the solvent d3 with the weakest desorption force among the five solvents is supplied from the solvent supply port D3. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

The desorption force of the solvent flowing through the sections 4 and 5 in the above sub-step (A3-3) is preferably the same as the desorption force of the solvent flowing through the section 5 in the above sub-step (A3-4).

As an example of the above sub-step (A2-4), a sub-step for implementing the following sub-step (A2-4ex) can be listed, but the above sub-step (A2-4) is not limited to the sub-step (A2-4ex). <Sub-step (A2-4ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the five solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the five solvents is supplied from the solvent supply port D2. The upstream end of the interval 4 is used as the solvent supply port D4, and the solvent d4 with the fourth strongest desorption force among the five solvents is supplied from the solvent supply port D4. The weak adsorption part is extracted from the weak adsorption part extraction port A. The solvent supply port D1 in this sub-step (A2-4ex) is provided in the same pipe as the solvent supply port D2 in the above-mentioned sub-step (A1-4ex).

As an example of the above sub-step (A3-4), a sub-step for implementing the following sub-step (A3-4ex) can be listed, but the above sub-step (A3-4) is not limited to the sub-step (A3-4ex). <Sub-step (A3-4ex)> The solvent d1 is supplied from the solvent supply port D1, and the strongly adsorbent portion is extracted from the strongly adsorbent portion extraction port C. The solvent d2 is supplied from the solvent supply port D2 in the sub-step (A2-4ex), and the downstream end of the interval 4 is used as the medium adsorbent portion extraction port B, and the medium adsorbent portion is extracted from the extraction port B. The upstream end of the interval 5 is used as the solvent supply port D5, and the solvent d5 with the third strongest desorption force among the five solvents is supplied from the solvent supply port D5, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

An example of a flow chart of an embodiment in which the above step (A) is sequentially executed with the above sub-steps (A1-4ex), (A2-4ex) and (A3-4ex) is shown in FIG8 . In FIG8 , a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower. In addition, the embodiment shown in FIG8 has 7 unit packed towers, including 1 unit packed tower in section 1, 2 unit packed towers in section 2, 2 unit packed towers in section 3, 1 unit packed tower in section 4, and 1 unit packed tower in section 5. After the step (A) of sequentially performing the above sub-steps (A1-4ex), (A2-4ex) and (A3-4ex) is completed, the stock solution supply port F, the elution supply port D, the weakly adsorbable part extraction port A, the medium adsorbable part extraction port B and the strong adsorbable part extraction port C are moved downstream while maintaining their relative positional relationship through step (B). Then, a flow chart of the state of sequentially performing the above sub-steps (A1-4ex), (A2-4ex) and (A3-4ex) is shown in FIG9. The unit packed towers arranged in each zone shown in FIG8 are shifted downstream by one unit of the unit packed tower in FIG9. At this time, "starting from step (A) shown in FIG. 8 and then performing step (B)" is regarded as one set of steps. By performing seven sets of steps, we return to the state shown in FIG. 8 again.

-Implementation Sample 5- Implementation Sample 5 uses a circulation system having more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this implementation sample 5, the following sub-steps (A1-5), (A2-5) and (A3-5) are sequentially implemented as step (A).

<Sub-step (A1-5)> The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the elution supply port D-III, and the elution d-III is supplied from the elution supply port D-III. The downstream end of the interval 5 is used as the weakly adsorbable part extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the elution flowing through the interval 3 the strongest, and making the desorption force of the elution flowing through the intervals 4 and 5 weaker than the desorption force of the elution flowing through the interval 3.

<Sub-step (A2-5)> Use the upstream end of the interval 1 as the solvent supply port D-I, and supply the solvent d-I from the solvent supply port D-I; use the downstream end of the interval 1 as the strong adsorption part extraction port C, and extract the strong adsorption part from the extraction port C; use the upstream end of the interval 2 as the solvent supply port D-II, and supply the solvent d-II from the solvent supply port D-II The solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than that of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than that of the solvent flowing through the zones 2 and 3.

<Sub-step (A3-5)> Supply the solvent d-I from the solvent supply port D-I, extract the strong adsorption part from the extraction port C, supply the solvent d-II from the solvent supply port D-II, use the downstream end of the interval 4 as the medium adsorption part extraction port B, extract the medium adsorption part from the extraction port B, use the upstream end of the interval 5 as the solvent supply port D-IV, and The solvent supply port D-IV supplies the solvent d-IV, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the solvent flowing through the segment 1 have the strongest desorption force, making the solvent flowing through the segments 2, 3 and 4 have weaker desorption force than the solvent flowing through the segment 1, and making the solvent flowing through the segment 5 have weaker desorption force than the solvent flowing through the segments 2, 3 and 4. The desorption force of the solvent flowing through the segment 1 in the sub-step (A3-5) is preferably the same as the desorption force of the solvent flowing through the segment 1 in the above sub-step (A2-5).

As an example of the above-mentioned sub-step (A1-5), a sub-step of implementing the following sub-step (A1-5ex) can be listed, but the above-mentioned sub-step (A1-5) is not limited to the sub-step (A1-5ex). ＜Sub-step (A1-5ex)＞ The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the elution supply port D3, and the elution d3 with the weakest desorption force among the four elutions is supplied from the elution supply port D3. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above sub-step (A2-5), a sub-step for implementing the following sub-step (A2-5ex) can be listed, but the above sub-step (A2-5) is not limited to the sub-step (A2-5ex). <Sub-step (A2-5ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 2 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The solvent d3 is supplied from the solvent supply port D3, and the weak adsorption part is extracted from the weak adsorption part extraction port A.

As an example of the above-mentioned sub-step (A3-5), a sub-step of implementing the following sub-step (A3-5ex) can be listed, but the above-mentioned sub-step (A3-5) is not limited to sub-step (A3-5ex). ＜Sub-step (A3-5ex)＞ The solvent d1 is supplied from the solvent supply port D1, the strongly adsorbent portion is extracted from the strongly adsorbent portion extraction port C, the solvent d2 is supplied from the solvent supply port D2, the downstream end of the interval 4 is used as the medium adsorbent portion extraction port B, the medium adsorbent portion is extracted from the extraction port B, the upstream end of the interval 5 is used as the solvent supply port D4, the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the embodiment in which each zone has a unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-5ex), (A2-5ex) and (A3-5ex) is shown in FIG13. In FIG13, a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower. After the step (A) of sequentially performing the above sub-steps (A1-5ex), (A2-5ex) and (A3-5ex) is completed, the stock solution supply port F, the elution supply port D, the weakly adsorbable part extraction port A, the medium adsorbable part extraction port B and the strong adsorbable part extraction port C are moved to the downstream side while maintaining their relative positional relationship through step (B). Then, the flow chart of the state of sequentially performing the above sub-steps (A1-5ex), (A2-5ex) and (A3-5ex) is shown in FIG14. The unit packed towers arranged in each compartment shown in FIG13 are displaced one by one to the downstream side in FIG14. At this time, "starting from step (A) shown in FIG. 13 and then performing step (B)" is regarded as one set of steps. By performing five sets of steps, we will return to the state shown in FIG. 13 again.

-Implementation Sample 6- Implementation Sample 6 uses a circulation system having more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this implementation sample 6, the following sub-steps (A1-6), (A2-6) and (A3-6) are sequentially implemented as step (A).

<Sub-step (A1-6)> The upstream end of the interval 1 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The downstream end of the interval 3 is used as the medium adsorption part extraction port B, and the medium adsorption part is extracted from the extraction port B. The upstream end of the interval 4 is used as the solvent supply port D-IV, and the solvent d-IV is supplied from the solvent supply port D-IV. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A. In this way, the desorption force of the solvent flowing through the intervals 1, 2 and 3 is the strongest, and the desorption force of the solvent flowing through the intervals 4 and 5 is weaker than the desorption force of the solvent flowing through the intervals 1, 2 and 3.

<Sub-step (A2-6)> The upstream end of the interval 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent The solvent supply port D-III is used to supply the solvent d-III from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zone 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3. The desorption force of the solvent flowing through the zone 1 in the sub-step (A2-6) is preferably stronger than the desorption force of the solvent flowing through the zone 1 in the above sub-step (A1-6). The solvent supply port D-I in the sub-step (A2-6) is provided in the same pipe as the solvent supply port D-II in the above-mentioned sub-step (A1-6).

<Sub-step (A3-6)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the upstream end of the zone 2 is used as the solvent supply port D-II, the solvent d-II is supplied from the solvent supply port D-II, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3. The desorption force of the solvent flowing through the interval 1 in the sub-step (A3-6) is preferably the same as the desorption force of the solvent flowing through the interval 1 in the above sub-step (A2-6).

As an example of the above sub-step (A1-6), a sub-step for implementing the following sub-step (A1-6ex) can be listed, but the above sub-step (A1-6) is not limited to the sub-step (A1-6ex). <Sub-step (A1-6ex)> The upstream end of the interval 1 is used as the solvent supply port D2, and the solvent d2 with the second strongest desorption force among the four solvents is supplied from the solvent supply port D2. The downstream end of the interval 3 is used as the medium adsorption part extraction port B, and the medium adsorption part is extracted from the extraction port B. The upstream end of the interval 4 is used as the solvent supply port D4, and the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above sub-step (A2-6), a sub-step for implementing the following sub-step (A2-6ex) can be listed, but the above sub-step (A2-6) is not limited to the sub-step (A2-6ex). <Sub-step (A2-6ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D3, and the solvent d3 with the weakest desorption force among the four solvents is supplied from the solvent supply port D3, and the weak adsorption part is extracted from the weak adsorption part extraction port A.

As an example of the above-mentioned sub-step (A3-6), a sub-step of implementing the following sub-step (A3-6ex) can be listed, but the above-mentioned sub-step (A3-6) is not limited to sub-step (A3-6ex). ＜Sub-step (A3-6ex)＞ The elution liquid d1 is supplied from the elution liquid supply port D1, and the strongly adsorbent part is extracted from the strongly adsorbent part extraction port C. The upstream end of the interval 2 is used as the elution liquid supply port D2, and the elution liquid d2 is supplied from the elution liquid supply port D2, and the elution liquid d3 is supplied from the elution liquid supply port D3, and the weakly adsorbent part is extracted from the weakly adsorbent part extraction port A.

Taking the embodiment in which each zone has one unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-6ex), (A2-6ex) and (A3-6ex) is shown in FIG15. In FIG15, a square frame represents one unit packed tower, and the number in the frame represents the number of the unit packed tower. After the step (A) of sequentially performing the above sub-steps (A1-6ex), (A2-6ex) and (A3-6ex) is completed, the stock solution supply port F, the elution supply port D, the weakly adsorbable part extraction port A, the medium adsorbable part extraction port B and the strong adsorbable part extraction port C are moved to the downstream side while maintaining their relative positional relationship through step (B), and then, the flow chart of the state of sequentially performing the above sub-steps (A1-6ex), (A2-6ex) and (A3-6ex) is shown in FIG16. The unit packed towers arranged in each compartment shown in FIG15 are displaced one by one to the downstream side in FIG16. At this time, "starting from step (A) shown in FIG. 15 and then performing step (B)" is regarded as one set of steps. By performing five sets of steps, we return to the state shown in FIG. 15.

-Implementation Sample 7- Implementation Sample 7 uses a circulation system having more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower." In addition, 4 types of eluents d-I to d-IV with different desorption forces are used as eluents. In this implementation sample 7, the following sub-steps (A1-7), (A2-7) and (A3-7) are sequentially implemented as step (A).

<Sub-step (A1-7)> Use the upstream end of the interval 1 as the solvent supply port D-I, supply the solvent d-I from the solvent supply port D-I, use the downstream end of the interval 1 as the strong adsorption part extraction port C, extract the strong adsorption part from the extraction port C, use the upstream end of the interval 3 as the stock solution supply port F, supply the stock solution from the stock solution supply port F, use the upstream end of the interval 4 as the solvent supply port D-III , the solvent d-III is supplied from the solvent supply port D-III, and the downstream side end of the zone 5 is used as the extraction port A. The weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zone 3 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3.

<Sub-step (A2-7)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent portion is extracted from the extraction port C, the upstream end of the zone 2 is used as the solvent supply port D-II, the solvent d-II is supplied from the solvent supply port D-II, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent portion is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3.

<Sub-step (A3-7)> Supply the solvent d-I from the solvent supply port D-I, extract the strong adsorption part from the extraction port C, supply the solvent d-II from the solvent supply port D-II, use the downstream end of the interval 4 as the medium adsorption part extraction port B, extract the medium adsorption part from the extraction port B, use the upstream end of the interval 5 as the solvent supply port D-IV, and The solvent supply port D-IV supplies the solvent d-IV, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4.

As an example of the above sub-step (A1-7), a sub-step for implementing the following sub-step (A1-7ex) can be listed, but the above sub-step (A1-7) is not limited to the sub-step (A1-7ex). <Sub-step (A1-7ex)> The upstream end of the interval 1 is used as the solvent supply port D1, and the solvent d1 with the strongest desorption force among the four solvents is supplied from the solvent supply port D1. The downstream end of the interval 1 is used as the strong adsorption part extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream end of the interval 4 is used as the solvent supply port D3, and the solvent d3 with the weakest desorption force among the four solvents is supplied from the solvent supply port D3. The downstream end of the interval 5 is used as the weak adsorption part extraction port A, and the weak adsorption part is extracted from the extraction port A.

As an example of the above-mentioned sub-step (A2-7), a sub-step of implementing the following sub-step (A2-7ex) can be listed, but the above-mentioned sub-step (A2-7) is not limited to the sub-step (A2-7ex). ＜Sub-step (A2-7ex)＞ The elution liquid d1 is supplied from the elution liquid supply port D1, the strongly adsorbent part is extracted from the strongly adsorbent part extraction port C, the elution liquid d3 is supplied from the elution liquid supply port D3, and the weakly adsorbent part is extracted from the weakly adsorbent part extraction port A.

As an example of the above-mentioned sub-step (A3-7), a sub-step of implementing the following sub-step (A3-7ex) can be listed, but the above-mentioned sub-step (A3-7) is not limited to sub-step (A3-7ex). <Sub-step (A3-7ex)> The solvent d1 is supplied from the solvent supply port D1, the strongly adsorbed part is extracted from the strongly adsorbed part extraction port C, the solvent d2 is supplied from the solvent supply port D2, the downstream end of the interval 4 is used as the medium adsorbed part extraction port B, the medium adsorbed part is extracted from the extraction port B, the upstream end of the interval 5 is used as the solvent supply port D4, the solvent d4 with the third strongest desorption force among the four solvents is supplied from the solvent supply port D4, and the weakly adsorbed part is extracted from the weakly adsorbed part extraction port A.

Taking the embodiment in which each zone has a unit packed tower as an example, a flow chart of the embodiment in which the above step (A) sequentially performs the above sub-steps (A1-7ex), (A2-7ex) and (A3-7ex) is shown in FIG17. In FIG17, a square frame represents a unit packed tower, and the number in the frame represents the number of the unit packed tower. After the step (A) of sequentially performing the above sub-steps (A1-7ex), (A2-7ex) and (A3-7ex) is completed, the stock solution supply port F, the elution supply port D, the weakly adsorbable part extraction port A, the medium adsorbable part extraction port B and the strong adsorbable part extraction port C are moved to the downstream side while maintaining their relative positional relationship through step (B), and then, the flow chart of the state of sequentially performing the above sub-steps (A1-7ex), (A2-7ex) and (A3-7ex) is shown in Figure 18. The unit packed towers arranged in each compartment shown in Figure 17 are displaced one by one to the downstream side in Figure 18. At this time, "starting from step (A) shown in FIG. 17 and then performing step (B)" is regarded as one set of steps. By performing five sets of steps, we will return to the state shown in FIG. 17.

In the method of the present invention, the target liquid is supplied to the target location, or the target liquid is extracted from the target location, by appropriately adjusting the operation of the pumps installed in various parts of the circulation system, and the opening and closing of the valves in various parts. That is, the method of supplying the target fluid in the circulation system or extracting the target part is known. In addition, the supply flow rate or extraction flow rate of each liquid can also be appropriately set according to the purpose of processing efficiency, etc.

In the method of the present invention, the component to be purified may be any one of a strongly adsorbable component, a moderately adsorbable component, and a weakly adsorbable component, but the method is more suitable for purifying the moderately adsorbable component. The method of the present invention can be applied to the purification of proteins. Since the method of the present invention can obtain a high-purity moderately adsorbable component, it is a method suitable for obtaining a high-purity target protein from a stock solution that contains not only the target protein but also its decomposition products or aggregates. The above-mentioned protein is not particularly limited, for example, an antibody can be the component to be purified. In the present invention, the so-called "antibody" may be a naturally occurring antibody, a chimeric antibody, or an antibody fragmented by an enzyme, etc. [for example, F(ab') ₂ fragment, Fab' fragment, Fab fragment]. In addition, it also includes single-chain antibodies or their two-chain antibodies (diabody) or three-chain antibodies (triabody), or microantibodies. In addition, it can also be a single domain antibody. In addition, these substances are only examples, and proteins and even their derivatives that have specific binding forces with respect to antigens are all included in the concept of antibodies of the present invention. In the method of the present invention, highly purified antibodies can also be used as antibody drugs. That is, by applying the method of the present invention to extract the antibodies contained in the stock solution, a method for producing an antibody drug can be provided. More specifically, using the method of the present invention, the culture medium of antibody-producing cells and/or the extract of antibody-producing cells are used as the stock solution, and the antibodies contained therein are extracted to obtain antibody drugs. In the present invention, the term "culture medium of antibody-producing cells" or "extract of antibody-producing cells" means a culture medium of antibody-producing cells or an extract of antibody-producing cells that has been subjected to various treatments such as centrifugation or chromatographic separation and is in a state of separation or purification to a certain extent.

In the method of the present invention, the adsorbent filled in the unit packing tower is appropriately selected according to the component to be purified, and various adsorbents can be used. For example, strongly acidic cation exchange resins, weakly acidic cation exchange resins, strongly alkaline anion exchange resins, weakly alkaline anion exchange resins, synthetic adsorbents, zeolites, silica gels, and functional group bonded silica gels (preferably octadecylsilane bonded silica gels). In addition, other gel filtration materials and affinity adsorption materials can be used as adsorbents. When the component to be purified is protein, the adsorbent is preferably an ion exchange resin. Among them, cation exchange resins are more suitable.

The simulated moving layer method layer analysis and separation system of the present invention is a system for implementing the method of the present invention. That is, the simulated moving layer method layer analysis and separation system of the present invention has the structure of the above-mentioned circulation system, and the circulation system is a system that can repeat the action of the above-mentioned step (A) and the action of the step (B) in sequence. [Implementation Example]

The present invention is further described below in detail based on embodiments, but the present invention is not limited to the following embodiments.

[Preparation of stock solution] Cells producing human immunoglobulin G2 (IgG2) are cultured, and the supernatant of the culture solution is desalted by dialysis, and the supernatant is adjusted to a salt concentration by adding NaCl as a stock solution. The content of the antibody, its fragments, and aggregates contained in the supernatant is as follows. In the table below, fragment 1 is a protein contained in the portion with a peak molecular weight of around 5,000 and a molecular weight of less than 25,000, and fragment 2 is a protein contained in the portion with a molecular weight of more than 25,000 and less than 50,000. In addition, the antibody is a protein contained in the portion with a peak molecular weight of around 150,000 and a molecular weight of more than 50,000 and less than 300,000. In addition, the aggregate is a protein contained in the portion with a molecular weight of more than 300,000. The following component composition was determined by high performance liquid chromatography (HPLC) using an analytical column (Tosoh TSKgel G3000SWXL).

[Table 1] Table 1 Components of the supernatant from the culture medium Concentration (g/L) Clip 1 0.02 Clip 2 0.326 antibody 1.564 Aggregate 0.090

[Adsorbent for a unit packed tower (column)] A cation exchange resin [trade name: Fractogel (registered trademark) EMD SO ₃ -(M), manufactured by Merck] was used as an adsorbent.

[Eluent] Phosphate buffer solutions with various NaCl concentrations were prepared using the following solutions A and B and used as eluents. ＜Solution A＞ 20mM phosphate buffer solution, pH 6.0. ＜Solution B＞ 20mM phosphate buffer solution containing 0.3M (17.53g/L) NaCl, pH 6.0.

[Comparison Example 1] Single column, step gradient <Column> Diameter 10 mm × length 100 mm, 1 column. <Stock solution> The NaCl concentration in the stock solution is 2.05 g/L. <Eluent> The following eluent was used.

[Table 2] Table 2 Type of solvent NaCl concentration of the elution solution (g/L) D1 2.39 D2 2.66 D3 17.53

<Operation conditions> The following steps 1 to 6 are performed in order. The flowchart of steps 1 to 6 is shown in FIG10. The following operation conditions are conditions that the recovery rate of fragments 1 and 2 in the weakly adsorbable part is more than 98%, the recovery rate of antibodies in the medium adsorbable part is more than 98%, and the recovery rate of aggregates in the strongly adsorbable part is more than 98%. This is also the same in each comparative example and embodiment described below.

[Table 3] Table 3 Steps Time (minutes) Flow rate (mL/min) 1 5.03 9.11 2 8.50 9.11 3 0.71 9.11 4 39.46 9.11 5 0.66 9.11 6 3.60 9.11

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed fraction, the recovery rate of antibodies in the medium-adsorbed fraction, and the recovery rate of aggregates in the strongly adsorbed fraction are shown in the table below. The recovery rate was calculated by 100×[mass in fraction]/[mass in stock solution].

[Table 4] Table 4 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.2 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation efficiency - The separation efficiency is the amount of stock solution treated (unit: "L (liter) - stock solution") per unit volume of adsorbent (unit: "L (liter) - R", R is the abbreviation of Resin) and per unit time (unit: "h (hour)"). In addition, in the multi-column system using multiple columns described later, the volume of adsorbent is the total amount of adsorbent contained in all columns. The separation efficiency in Comparative Example 1 is 6.04 (L - stock solution) / (L - R) h.

[Comparison Example 2] Single column, step gradient <Column> Diameter 10 mm × length 400 mm, 1 column. <Stock solution> The NaCl concentration in the stock solution is 2.05 g/L. <Eluent> The following eluent was used.

[Table 5] Table 5 Type of solvent NaCl concentration of the elution solution (g/L) D1 2.35 D2 2.62 D3 17.53

<Operation conditions> Perform the following steps 1 to 6 in order. The flow chart of steps 1 to 6 is shown in Figure 10.

[Table 6] Table 6 Steps Time (minutes) Flow rate (mL/min) 1 5.99 43.3 2 6.35 43.3 3 0.63 43.3 4 22.48 43.3 5 0.58 43.3 6 3.55 43.3

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 7] Table 7 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation efficiency - The separation efficiency in Comparative Example 2 is 12.51 (L-stock solution)/(L-R) h.

[Comparison Example 3] Multi-column, gradient, and simulated moving layer method <Column> Diameter 10 mm × length 100 mm, 4 columns. <Stock solution> The NaCl concentration in the stock solution is 2.23 g/L. <Eluent> The following eluent was used.

[Table 8] Table 8 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.28 D3 2.23

<Operation conditions> Figure 11 shows a flow chart of the operation of Comparative Example 3. The first to fourth steps shown in Figure 11 are taken as one cycle, and 10 cycles are implemented. Between each step, a step is implemented in which the stock solution supply port F, the solvent supply port D (D1 to D3), the weak adsorption part extraction port A, the medium adsorption part extraction port B, and the strong adsorption part extraction port C are moved downstream by one column while maintaining their relative positional relationship. In addition, the first to fourth steps shown in Figure 11 correspond to step (A) of the present invention, but unlike step (A), they are not composed of a plurality of sub-steps (in other words, one step is composed of one sub-step). "D1", "D2" and "D3" in Figure 11 are all solvent supply ports, which supply solvents D1, D2 and D3 respectively. "C" in Figure 11 is the strong adsorption part extraction port, which extracts the strong adsorption part. Similarly, "B" is the medium adsorption part extraction port, which extracts the medium adsorption part, and "A" is the weak adsorption part extraction port, which extracts the weak adsorption part. The supply flow rates of the solvent (D1~D3) and the stock solution (F) in each step shown in Figure 11 are as described below. In addition, the table below does not record the flow rate of the extracted liquid, but the flow rate of the strong adsorption part extracted from the strong adsorption part extraction port C is the same as the flow rate of the supply solvent D1. In addition, the flow rate of the medium adsorption part extracted from the medium adsorption part extraction port B is the same as the flow rate of the supply solvent D2. In addition, the flow rate of the weakly adsorbent part extracted from the weakly adsorbent part extraction port A is the sum of the flow rate of the supplied eluent D3 and the flow rate of the supplied stock solution from the stock solution supply port F. That is, the supply flow rate and the extraction flow rate are always the same, and this is also the case in the subsequent comparative examples and even the embodiment.

[Table 9] Table 9 Time for 1 step (min) Flow rate (mL/min) 29.9 D1 2.14 D2 18.7 D3 1.79 f/3.73

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 10] Table 10 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 More than 99 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Comparative Example 3 is 7.11 (L-stock solution)/(L-R) h.

[Comparison Example 4] Multi-column, gradient, and simulated moving layer method <Column> Diameter 10 mm × length 100 mm, 4 columns. <Stock solution> The NaCl concentration in the stock solution is 2.24 g/L. <Eluent> The following eluent was used.

[Table 11] Table 11 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.26 D3 2.24

<Operation conditions> Figure 12 shows a flow chart of the operation of Comparative Example 4. The first to fourth steps shown in Figure 12 are taken as one cycle, and 10 cycles are implemented. In addition, each step shown in Figure 12 is composed of two sub-steps, the first sub-step and the second sub-step. In the second sub-step, the supply and extraction of liquid are not performed, but the fluid in the circulation system is circulated. The supply flow rates of the elution solution (D1 to D3) and the stock solution (F) in each step shown in Figure 12 are as follows.

[Table 12] Table 12 Substep duration (min) Flow rate (mL/min) Sub-step 1 32.7 D1 1.90 D2 17.4 D3 1.33 f/3.82 Sub-step 2 0.48 Cycle 0.94

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 13] Table 13 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 More than 99 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Comparative Example 4 is 7.19 (L-stock solution)/(L-R) h.

[Example 1] Multi-column, gradient, simulated moving layer method <Column> Diameter 10mm×length 100mm, 4 columns. <Stock solution> The NaCl concentration in the stock solution is 1.93g/L. <Eluent> The following eluent was used.

[Table 14] Table 14 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 3.59 D3 1.93 D4 2.21

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in Figure 2. This step (A) and the subsequent step (B) are used as one set of steps, and 4 sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 15] Table 15 Substep duration (min) Flow rate (mL/min) Sub-step (A1-1) 4.93 D1 10.76 D2 4.41 f/21.77 Sub-step (A2-1) 1.73 D1 10.76 D2 4.41 D3 21.77 Sub-steps (A3-1) 3.75 D1 13.87 D2 5.62 D4 15.21

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 16] Table 16 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 1 is 19.696 (L-stock solution)/(L-R) h.

[Example 2] Multi-column, gradient, simulated moving layer method <Column> 4 columns, diameter 10 mm × length 100 mm. <Stock solution> The NaCl concentration in the stock solution was 2.02 g/L. <Eluent> The following eluent was used. [Table 17] Table 17 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 3.57 D3 2.02 D4 2.21

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in FIG4. This step (A) and the subsequent step (B) are considered as one set of steps, and four sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 18] Table 18 Substep duration (min) Flow rate (mL/min) Sub-steps (A1-2) 6.82 D2 2.39 f/17.65 Sub-step (A2-2) 1.67 D1 3.25 D2 2.39 D3 17.65 Sub-steps (A3-2) 3.86 D1 7.80 D2 5.38 D4 7.37

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 19] Table 19 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 2 is 18.610 (L-stock solution)/(L-R) h.

[Example 3] Multi-column, gradient, simulated moving layer method <Column> 10 mm diameter × 805 mm length, 5 columns. <Stock solution> The NaCl concentration in the stock solution was 2.46 g/L. <Eluent> The following eluent was used. [Table 20] Table 20 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.46 D3 0.00 D4 1.98

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in Figure 6. This step (A) and the subsequent step (B) are considered as one set of steps, and 5 sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 21] Table 21 Time for 1 step (min) Flow rate (mL/min) Sub-steps (A1-3) 10.39 D2 0.003 D3 4.56 f/18.54 Sub-steps (A2-3) 3.66 D1 2.90 D2 8.95 D3 2.20 Sub-steps (A3-3) 8.25 D1 2.49 D2 21.42 D4 2.03

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 22] Table 22 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 3 is 16.499 (L-stock solution)/(L-R) h.

[Example 4] Multi-column, gradient, simulated moving layer method <Column> 7 columns, diameter 10 mm × length 100 mm. <Stock solution> The NaCl concentration in the stock solution was 2.57 g/L. <Eluent> The following eluent was used. [Table 23] Table 23 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.57 D3 0.00 D4 0.37 D5 1.80

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in FIG8. This step (A) and the subsequent step (B) are considered as one set of steps, and 7 sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D5) and the stock solution (F) in each step (A) are as follows.

[Table 24] Table 24 Time for 1 step (min) Flow rate (mL/min) Sub-steps (A1-4) 8.18 D2 0.004 D3 6.86 f/16.07 Sub-steps (A2-4) 3.84 D1 2.94 D2 10.68 D4 5.73 Sub-steps (A3-4) 4.47 D1 3.43 D2 19.02 D5 2.18

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 25] Table 25 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 4 is 15.225 (L-stock solution)/(L-R) h.

As described above, it can be seen that in the simulated shifting layer separation, by using two or more eluents and making the positional relationship of the weakly adsorbable fraction extraction port A, the medium adsorbable fraction extraction port B, the strongly adsorbable fraction extraction port C, and the stock solution supply port F in the circulation system the specific relationship defined by the present invention, the weakly adsorbable component, the medium adsorbable component, and the strongly adsorbable component can be extracted with sufficient high purity with a smaller amount of adsorbent. This embodiment discloses the technical content of obtaining the target antibody in the medium adsorbable fraction with high purity and high efficiency.

[Example 5] Multi-column, gradient, simulated moving layer method <Column> Diameter 10mm×length 80mm, 5 columns. <Stock solution> The NaCl concentration in the stock solution is 2.46g/L. <Eluent> The following eluent was used.

[Table 26] Table 26 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.46 D3 0.00 D4 1.98

The combination of sub-steps shown in FIG13 constitutes step (A). This step (A) and the subsequent step (B) are considered as one set of steps, and five sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 27] Table 27 Time for 1 step (min) Flow rate (mL/min) Sub-steps (A1-5) 10.39 D3 4.56 f/18.54 Sub-steps (A2-5) 3.66 D1 2.90 D2 8.95 D3 2.20 Sub-steps (A3-5) 8.25 D1 2.49 D2 21.42 D4 2.03

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 28] Table 28 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 5 is 16.502 (L-stock solution)/(L-R) h.

[Example 6] Multi-column, gradient, simulated moving layer method <Column> 10 mm diameter × 80 mm length, 5 columns. <Stock solution> The NaCl concentration in the stock solution was 2.55 g/L. <Eluent> The following eluent was used. [Table 29] Table 29 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.55 D3 0.00 D4 2.03

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in FIG15. This step (A) and the subsequent step (B) are considered as one set of steps, and 5 sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 30] Table 30 Time for 1 step (min) Flow rate (mL/min) Sub-steps (A1-6) 5.98 D2 25.06 D4 1.91 Sub-steps (A2-6) 8.24 D1 2.35 D3 5.30 f/21.05 Sub-steps (A3-6) 3.31 D1 2.92 D2 10.55 D3 2.66

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 31] Table 31 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 6 is 18.898 (L-stock solution)/(L-R) h.

[Example 7] Multi-column, gradient, simulated moving layer method <Column> 10 mm diameter × 80 mm length, 5 columns. <Stock solution> The NaCl concentration in the stock solution was 2.55 g/L. <Eluent> The following eluent was used. [Table 32] Table 32 Type of solvent NaCl concentration of the elution solution (g/L) D1 17.53 D2 2.55 D3 0.00 D4 2.03

<Operation conditions> Step (A) is composed of the combination of sub-steps shown in FIG17. This step (A) and the subsequent step (B) are considered as one set of steps, and 5 sets are performed as one cycle, and 10 cycles are performed. The supply flow rates of the elution solution (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 33] Table 33 Time for 1 step (min) Flow rate (mL/min) Sub-steps (A1-7) 10.39 D1 2.90 D3 4.56 f/18.54 Sub-steps (A2-7) 3.66 D1 2.90 D2 8.95 D3 2.20 Sub-steps (A3-7) 8.25 D1 2.49 D2 21.42 D4 2.03

<Results> -Recovery rate- The recovery rates of fragments 1 and 2 in the weakly adsorbed part, the recovery rate of antibodies in the medium adsorbed part, and the recovery rate of aggregates in the strongly adsorbed part are shown in the table below.

[Table 34] Table 34 part Recycled Content Recovery rate(%) Weakly adsorbable part Clip 1 More than 99 Clip 2 98.0 Medium adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

- Separation treatment efficiency - The separation treatment efficiency in Example 7 is 16.502 (L-stock solution)/(L-R) h.

Although the present invention is described together with its implementation modes, unless otherwise expressly stated, the present invention is not limited to a certain detail in the description, but should be broadly interpreted without violating the spirit and scope of the invention disclosed in the attached claims.

This case claims priority based on Patent Application No. 2018-215950 filed in Japan on November 16, 2018 and Patent Application No. 2019-088523 filed in Japan on May 8, 2019, and the contents of these cases are incorporated herein by reference as part of the description of this specification.

111: Piping 1~7: Unit packed tower number 2a,2b,2c,2d: Weak adsorption part extraction pipeline 2J: Weak adsorption part confluence pipe 3a,3b,3c,3d: Medium adsorption part extraction pipeline 3J: Medium adsorption part confluence pipe 4a,4b,4c,4d: Strong adsorption part extraction pipeline 4J: Strong adsorption part confluence pipe 116: Raw liquid tank 117: Raw liquid 8a,8b,8c,8d: Eluent tank 9a,9b,9c,9d: Eluent 10a,10b,10c,10d: Unit packed tower (column) 11a,11b, 11c, 11d: Raw liquid supply branch pipeline 11: Raw liquid supply pipeline 12, 13, 14, 15: Eluate supply pipeline 12a, 12b, 12c, 12d: Eluate supply branch pipeline 13a, 13b, 13c, 13d: Eluate supply branch pipeline 14a, 14b, 14c, 14d: Eluate supply branch pipeline 15a, 15b, 15c, 15d: Eluate supply branch pipeline 100: Circulation system (A): Steps (A1-1), (A2-1), (A3-1), (A1-2), (A2-2), (A3-2), (A 1-3), (A2-3), (A3-3), (A1-4), (A2-4), (A3-4), (A1-5), (A2-5), (A3-5), (A1-6), (A2-6), (A3-6), (A1-7), (A2-7), (A3-7): Sub-steps A1, A2, A3, A4: Weak adsorption partial extraction valve Ab: Adsorbent A: Weak adsorption partial extraction outlet B1, B2, B3, B4: Medium adsorption partial extraction valve B: Medium adsorption partial extraction outlet C1, C2, C3, C4: Strong adsorption partial extraction valve C: Strong adsorption partial extraction valve Extraction outlet D1, D2, D3, D4, D5: Solvent supply port E1a, E2a, E3a, E4a: Solvent supply valve E1b, E2b, E3b, E4b: Solvent supply valve E1c, E2c, E3c, E4c: Solvent supply valve E1d, E2d, E3d, E4d: Solvent supply valve F1, F2, F3, F4: Raw liquid supply valve F: Raw liquid supply port P1: Circulation pump P2: Raw liquid supply pump P3, P4, P5, P6: Solvent supply pump R1, R2, R3, R4: Block valve T1, T2, T3, T4: Check valve

[FIG. 1] is a system diagram showing an example of the simulated moving layer method analysis and separation system of the present invention. [FIG. 2] is a flowchart of each sub-step constituting step (A) in one embodiment of the simulated moving layer method analysis and separation method of the present invention. [FIG. 3] is a flowchart of each sub-step constituting step (A) after completing step (A) shown in FIG. 2 and implementing step (B). [FIG. 4] is a flowchart of each sub-step constituting step (A) in another embodiment of the simulated moving layer method analysis and separation method of the present invention. [Figure 5] is a flowchart of each sub-step of step (A) after completing step (A) shown in Figure 4 and implementing step (B). [Figure 6] is a flowchart of each sub-step of step (A) in another embodiment of the simulated moving layer analysis and separation method of the present invention. [Figure 7] is a flowchart of each sub-step of step (A) after completing step (A) shown in Figure 6 and implementing step (B). [Figure 8] is a flowchart of each sub-step of step (A) in another embodiment of the simulated moving layer analysis and separation method of the present invention. [Fig. 9] is a flowchart of each sub-step constituting step (A) after completing step (A) shown in Fig. 8 and implementing step (B). [Fig. 10] is a flowchart showing the operation step of the single column and step gradient in Comparative Examples 1 and 2. [Fig. 11] is a flowchart showing the operation step of the simulated moving layer method analysis and separation in Comparative Example 3. [Fig. 12] is a flowchart showing the operation step of the simulated moving layer method analysis and separation in Comparative Example 4. [Fig. 13] is a flowchart showing each sub-step constituting step (A) in yet another embodiment of the simulated moving layer method analysis and separation method of the present invention. [Figure 14] is a flowchart of each sub-step of step (A) after completing step (A) shown in Figure 13 and implementing step (B). [Figure 15] is a flowchart of each sub-step of step (A) in another embodiment of the simulated moving layer analysis and separation method of the present invention. [Figure 16] is a flowchart of each sub-step of step (A) after completing step (A) shown in Figure 15 and implementing step (B). [Figure 17] is a flowchart of each sub-step of step (A) in another embodiment of the simulated moving layer analysis and separation method of the present invention. [Fig. 18] is a flow chart of the sub-steps constituting step (A) after completing step (A) shown in Fig. 17 and implementing step (B).

111: Piping

2a, 2b, 2c, 2d: Weakly adsorbable part extraction pipeline

2J: Weakly adsorbable partial confluence tube

3a,3b,3c,3d: Extraction pipeline for the medium adsorption part

3J: Medium adsorption part confluence pipe

4a, 4b, 4c, 4d: Strong adsorption part extraction pipeline

4J: Strong adsorption partial confluence pipe

116: Original liquid tank

117: stock solution

8a,8b,8c,8d: Dissolution tank

9a,9b,9c,9d: Dissolution liquid

10a, 10b, 10c, 10d: Unit packed tower (column)

11a, 11b, 11c, 11d: Raw liquid supply branch pipeline

11: Raw liquid supply pipeline

12,13,14,15: Dissolution liquid supply pipeline

12a, 12b, 12c, 12d: branch pipelines for dissolution supply

13a, 13b, 13c, 13d: branch pipelines for dissolving liquid

14a, 14b, 14c, 14d: branch pipelines for dissolving liquid

15a, 15b, 15c, 15d: branch pipelines for dissolving liquid

100: Circulatory system

A1, A2, A3, A4: Weak adsorption partial extraction valve

Ab: adsorbent

B1, B2, B3, B4: Medium adsorption partial extraction valve

C1, C2, C3, C4: Strong adsorption partial extraction valve

E1a, E2a, E3a, E4a: Solvent supply valve

E1b, E2b, E3b, E4b: Solvent supply valve

E1c, E2c, E3c, E4c: Solvent supply valve

E1d, E2d, E3d, E4d: Solvent supply valve

F1, F2, F3, F4: Raw liquid supply valve

P1: Circulation pump

P2: Raw liquid supply pump

P3, P4, P5, P6: solution supply pump

R1, R2, R3, R4: Blocking valve

T1, T2, T3, T4: Check valve

Claims (13)

A stratification separation method using a simulated moving layer method comprises a circulation system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly through piping, and the steps of separating a weakly adsorbable component, a strongly adsorbable component, and a medium adsorbable component with an adsorption between the two components contained in a stock solution by using two or more solvents; the stratification separation method using a simulated moving layer method is characterized in that: a stock solution supply port F is provided in the piping of the circulation system, and a supply port F corresponding to each of the two or more solvents is provided. The invention relates to a method for preparing a liquid phase separation tower having two or more solvent supply ports D, an extraction port A for a weakly adsorbable portion containing the weakly adsorbable component, an extraction port B for a medium adsorbable portion containing the medium adsorbable component, and an extraction port C for a strong adsorbable portion containing the strong adsorbable component; and the positions of the stock solution supply port F, the extraction port A, the extraction port B, and the extraction port C are set as follows (a) to (c): (a) the extraction port B is set on the downstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (b) the extraction port C is set (c) the extraction port A is disposed in a pipe having the raw liquid supply port F, or the extraction port C is disposed on the upstream side of the raw liquid supply port F and sandwiching at least one unit packed tower; (c) the extraction port A is disposed in a pipe having the extraction port B, or the extraction port A is disposed on the downstream side of the extraction port B and sandwiching at least one unit packed tower; the layer separation method is a simulated moving layer layer separation method including sequentially repeating the following steps (A) and (B): [Step (A)] respectively or separately from The step of supplying the stock solution from the stock solution supply port F, supplying two or more types of solvents from the two or more solvent supply ports D, and extracting the weakly adsorbable part from the extraction port A, the medium adsorbable part from the extraction port B, and the strongly adsorbable part from the extraction port C simultaneously or separately; [Step (B)] After the step (A) is completed, the step of moving the stock solution supply port F, the solvent supply port D, the extraction port A, the extraction port B, and the extraction port C to the downstream side while maintaining their relative positional relationship. As in claim 1, the simulated moving layer analysis and separation method, wherein the step (A) is composed of a plurality of sub-steps; the plurality of sub-steps include: a sub-step of supplying the stock solution; and a sub-step of not supplying the stock solution. The simulated moving layer separation method of claim 1 or 2, wherein the extraction port C is set at the downstream side of the solvent supply port D1 for supplying the solvent d1 with the strongest desorption force among two or more solvents; at least one unit packed tower is arranged between the solvent supply port D1 and the extraction port C; in the step (A), during the supply of the solvent d1, the same amount of the strongly adsorbent part as the supply amount of the solvent d1 is extracted from the extraction port C. The simulated moving layer separation method of claim 1 or 2, wherein the extraction port B is set at the downstream side of the solvent supply port D2 for supplying the solvent d2 with the second strongest desorption force among two or more solvents; at least one unit packed tower is arranged between the solvent supply port D2 and the extraction port B; In the step (A), a time band is set for extracting the same amount of the medium adsorbent part as the supply amount of the solvent d2 from the extraction port B during the supply of the solvent d2. A simulated moving layer separation method as claimed in claim 1 or 2, wherein 4 to 6 solvents with different desorption forces are used. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 4 unit packed towers, and the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-1), (A2-1) and (A3-1) are performed in the step (A): <Sub-step (A1-1)> The upstream end of the section 1 is used as the eluting liquid supply port D-I, and the eluting liquid d-I is supplied from the eluting liquid supply port D-I, and the downstream end of the section 1 is used as the extraction port C, and the strong suction liquid is extracted from the extraction port C. The adsorption part, the upstream end of the interval 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II, the upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F, and the downstream end of the interval 4 is used as the extraction port A, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the interval 1 the strongest, making the desorption force of the solvent flowing through the interval 2 weaker than the desorption force of the solvent flowing through the interval 1, and making the desorption force of the solvent flowing through the intervals 3 and 4 weaker than the desorption force of the solvent flowing through the interval 2; <Sub-step (A2-1)> From the solvent supply port D-I Supply the solvent d-I, extract the strongly adsorbent part from the extraction port C, supply the solvent d-II from the solvent supply port D-II, use the upstream end of the zone 3 as the solvent supply port D-III, supply the solvent d-III from the solvent supply port D-III, and extract the weakly adsorbent part from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2; <Sub-step (A3-1)> Supply the solvent from the solvent supply port D-I d-I, extract the strong adsorption part from the extraction port C, supply the solvent d-II from the solvent supply port D-II, use the downstream end of the zone 3 as the extraction port B, extract the medium adsorption part from the extraction port B, use the upstream end of the zone 4 as the solvent supply port D-IV, supply the solvent d-IV from the solvent supply port D-IV, and extract the weak adsorption part from the extraction port A, thereby making the solvent flowing through the zone 1 have the strongest desorption force, making the solvent flowing through the zones 2 and 3 have a weaker desorption force than the solvent flowing through the zone 1, and making the solvent flowing through the zone 4 have a weaker desorption force than the solvent flowing through the zones 2 and 3. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 4 unit packed towers, and the circulation system is divided into 4 sections 1 to 4 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-2), (A2-2) and (A3-2) are performed in the step (A): <Sub-step (A1-2)> The upstream end of section 1 is used as the eluting liquid supply port D-II, and the eluting liquid d-II is supplied from the eluting liquid supply port D-II, and the upstream end of section 3 is used as the raw liquid supply port D-II. The raw liquid is supplied from the raw liquid supply port F, and the downstream end of the zone 4 is used as the extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, and making the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zones 1 and 2; <Sub-step (A2-2)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-I is supplied from the solvent supply port D-I, the downstream end of the zone 1 is used as the extraction port C, and the strongly adsorbable part is extracted from the extraction port C, and the upstream end of the zone 2 is used as the solvent supply port D-II, The solvent d-II is supplied from the solvent supply port D-II, and the upstream end of the zone 3 is used as the solvent supply port D-III. The solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbed part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zone 2 weaker than the desorption force of the solvent flowing through the zone 1, and the desorption force of the solvent flowing through the zones 3 and 4 weaker than the desorption force of the solvent flowing through the zone 2; <Sub-step (A3-2)> The solvent d-I is supplied from the solvent supply port D-I, and the strongly adsorbed part is extracted from the extraction port C, The solvent d-II is supplied from the solvent supply port D-II in sub-step (A2-2), the downstream end of the zone 3 is used as the extraction port B, and the medium adsorption part is extracted from the extraction port B. The upstream end of the zone 4 is used as the solvent supply port D-IV, and the solvent d-IV is supplied from the solvent supply port D-IV, and the weak adsorption part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zone 4 weaker than the desorption force of the solvent flowing through the zones 2 and 3. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 5 unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least one unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-3), (A2-3) and (A3-3) are performed in the step (A): <Sub-step (A1-3)> The upstream end of section 1 is used as the eluting liquid supply port D-II, and the eluting liquid d-II is supplied from the eluting liquid supply port D-II, the upstream end of section 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F, and the upstream end of section 4 is used as the stock solution supply port F. The downstream end of the zone 5 is used as the solvent supply port D-III, and the solvent d-III is supplied from the solvent supply port D-III. The downstream end of the zone 5 is used as the extraction port A, and the weakly adsorbed part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, making the desorption force of the solvent flowing through the zone 3 the same as or weaker than the desorption force of the solvent flowing through the zones 1 and 2, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A2-3)> The upstream end of the zone 1 is used as the solvent supply port D-I, and the solvent d-III is supplied from the solvent supply port D-I. I, take the downstream end of zone 1 as the extraction port C, extract the strongly adsorbent part from the extraction port C, take the upstream end of zone 2 as the solvent supply port D-II, supply the solvent d-II from the solvent supply port D-II, supply the solvent d-III from the solvent supply port D-III, and extract the weakly adsorbent part from the extraction port A, thereby making the desorption force of the solvent flowing through zone 1 the strongest, making the desorption force of the solvent flowing through zones 2 and 3 weaker than the desorption force of the solvent flowing through zones 1, and making the desorption force of the solvent flowing through zones 4 and 5 weaker than the desorption force of the solvent flowing through zones 2 and 3; <Sub-step (A3-3)> supplying the solvent from the solvent supply port D-I d-I, extract the strong adsorption part from the extraction port C, supply the solvent d-II from the solvent supply port D-II in sub-step (A2-3), use the downstream end of the interval 4 as the extraction port B, extract the medium adsorption part from the extraction port B, use the upstream end of the interval 5 as the solvent supply port D-IV, supply the solvent d-IV from the solvent supply port D-IV, and extract the weak adsorption part from the extraction port A, thereby making the solvent flowing through interval 1 have the strongest desorption force, making the solvent flowing through intervals 2, 3 and 4 have a weaker desorption force than the solvent flowing through interval 1, and making the solvent flowing through interval 5 have a weaker desorption force than the solvent flowing through intervals 2, 3 and 4. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 7 unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-4), (A2-4) and (A3-4) are implemented in the step (A): <Sub-step (A1-4)> The upstream end of section 1 is used as the eluting liquid supply port D-II, and the eluting liquid d-II is supplied from the eluting liquid supply port D-II, the upstream end of section 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F, and the upstream end of section 4 is used as the stock solution supply port F. As the solvent supply port D-III, the solvent d-III is supplied from the solvent supply port D-III, and the downstream end of the zone 5 is used as the extraction port A, and the weakly adsorbed part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1 and 2 the strongest, making the desorption force of the solvent flowing through the zone 3 the same as or weaker than the desorption force of the solvent flowing through the zones 1 and 2, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A2-4)> Taking the upstream end of the zone 1 as the solvent supply port D-I, the solvent d-I is supplied from the solvent supply port D-I, and the downstream end of the zone 1 is used as the extraction port A. The side end of the zone 2 is used as the extraction port C, and the strongly adsorbent part is extracted from the extraction port C. The upstream side end of the zone 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II. The upstream side end of the zone 4 is used as the solvent supply port D-IV, and the solvent d-IV is supplied from the solvent supply port D-IV. The weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; <Sub-step (A3-4)> From the solvent supply port D -I supplies the solvent d-I, extracts the strong adsorption part from the extraction port C, supplies the solvent d-II from the solvent supply port D-II in the sub-step (A2-4), takes the downstream end of the interval 4 as the extraction port B, extracts the medium adsorption part from the extraction port B, takes the upstream end of the interval 5 as the solvent supply port D-V, supplies the solvent d-V from the solvent supply port D-V, and extracts the weak adsorption part from the extraction port A, thereby making the desorption force of the solvent flowing through the interval 1 the strongest, making the desorption force of the solvent flowing through the intervals 2, 3 and 4 weaker than the desorption force of the solvent flowing through the interval 1, and making the desorption force of the solvent flowing through the interval 5 weaker than the desorption force of the solvent flowing through the intervals 2, 3 and 4. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 5 unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-5), (A2-5) and (A3-5) are performed in the step (A): <Sub-step (A1-5)> The upstream end of section 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F, and the upstream end of section 4 is used as the eluting liquid supply port D-III , supplying solvent d-III from the solvent supply port D-III, taking the downstream end of the zone 5 as the extraction port A, and extracting the weakly adsorbable part from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 3 the strongest, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zone 3; <Sub-step (A2-5)> taking the upstream end of the zone 1 as the solvent supply port D-I, supplying solvent d-I from the solvent supply port D-I, taking the downstream end of the zone 1 as the extraction port C, and extracting the strongly adsorbable part from the extraction port C, taking the upstream end of the zone 2 as the solvent The solvent supply port D-II supplies the solvent d-II from the solvent supply port D-II, supplies the solvent d-III from the solvent supply port D-III, and extracts the weakly adsorbable portion from the extraction port A, thereby making the desorption force of the solvent flowing through the interval 1 the strongest, making the desorption force of the solvent flowing through the intervals 2 and 3 weaker than the desorption force of the solvent flowing through the interval 1, and making the desorption force of the solvent flowing through the intervals 4 and 5 weaker than the desorption force of the solvent flowing through the intervals 2 and 3; <Sub-step (A3-5)> supplying the solvent d-I from the solvent supply port D-I, and extracting the strongly adsorbable portion from the extraction port C, The solvent d-II is supplied from the solvent supply port D-II, the downstream end of the zone 4 is used as the extraction port B, and the medium adsorption part is extracted from the extraction port B. The upstream end of the zone 5 is used as the solvent supply port D-IV, and the solvent d-IV is supplied from the solvent supply port D-IV, and the weak adsorption part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2, 3 and 4 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zone 5 weaker than the desorption force of the solvent flowing through the zones 2, 3 and 4. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 5 unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-6), (A2-6) and (A3-6) are implemented in the step (A): <Sub-step (A1-6)> The upstream end of section 1 is used as the eluting liquid supply port D-II, and the eluting liquid d-II is supplied from the eluting liquid supply port D-II, and the downstream end of section 3 is used as the extraction port B , extract the medium adsorption part from the extraction port B, use the upstream end of the zone 4 as the solvent supply port D-IV, supply the solvent d-IV from the solvent supply port D-IV, use the downstream end of the zone 5 as the extraction port A, and extract the weak adsorption part from the extraction port A, thereby making the desorption force of the solvent flowing through the zones 1, 2 and 3 the strongest, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 1, 2 and 3; <Sub-step (A2-6)> Use the upstream end of the zone 1 as the solvent supply port D-I, supply the solvent d-I from the solvent supply port D-I, and use the downstream end of the zone 1 as the extraction port A. The side end of the zone 3 is used as the extraction port C, and the strong adsorption part is extracted from the extraction port C. The upstream side end of the zone 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F. The upstream side end of the zone 4 is used as the elution supply port D-III, and the elution d-III is supplied from the elution supply port D-III. The weak adsorption part is extracted from the extraction port A, thereby making the desorption force of the elution flowing through the zone 1 the strongest, making the desorption force of the elution flowing through the zone 3 weaker than the desorption force of the elution flowing through the zone 1, and making the desorption force of the elution flowing through the zones 4 and 5 weaker than the desorption force of the elution flowing through the zone 3; <sub-step (A3- 6)> The solvent d-I is supplied from the solvent supply port D-I, the strongly adsorbent part is extracted from the extraction port C, the upstream end of the zone 2 is used as the solvent supply port D-II, the solvent d-II is supplied from the solvent supply port D-II, the solvent d-III is supplied from the solvent supply port D-III, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, making the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zone 1, and making the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3. A simulated moving layer separation method as claimed in claim 1 or 2, wherein the circulation system has more than 5 unit packed towers, and the circulation system is divided into 5 sections 1 to 5 connected in a ring shape from the upstream side to the downstream side in a manner that each section has at least 1 unit packed tower. In addition, the two or more eluting liquids are used, and the following sub-steps (A1-7), (A2-7) and (A3-7) are implemented in the step (A): <Sub-step (A1-7)> The upstream end of section 1 is used as the eluting liquid supply port D-I, and the eluting liquid d-I is supplied from the eluting liquid supply port D-I, and the downstream end of section 1 is used as the extraction port C, and the strongly adsorbent is extracted from the extraction port C. The upstream end of the interval 3 is used as the stock solution supply port F, and the stock solution is supplied from the stock solution supply port F; the upstream end of the interval 4 is used as the elution supply port D-III, and the elution d-III is supplied from the elution supply port D-III; the downstream end of the interval 5 is used as the extraction port A, and the weakly adsorbent part is extracted from the extraction port A, thereby making the desorption force of the elution flowing through the interval 1 the strongest, making the desorption force of the elution flowing through the interval 3 weaker than the desorption force of the elution flowing through the interval 1, and making the desorption force of the elution flowing through the intervals 4 and 5 weaker than the desorption force of the elution flowing through the interval 3; <Sub-step (A2-7)> supplying the elution from the elution supply port D-I to the The solvent d-I is extracted from the extraction port C with a strong adsorption part, and the upstream end of the zone 2 is used as the solvent supply port D-II, and the solvent d-II is supplied from the solvent supply port D-II, and the solvent d-III is supplied from the solvent supply port D-III, and the weak adsorption part is extracted from the extraction port A, thereby making the desorption force of the solvent flowing through the zone 1 the strongest, the desorption force of the solvent flowing through the zones 2 and 3 weaker than the desorption force of the solvent flowing through the zones 1, and the desorption force of the solvent flowing through the zones 4 and 5 weaker than the desorption force of the solvent flowing through the zones 2 and 3; <Sub-step (A3-7)> supplying the solvent d-I from the solvent supply port D-I -I, extract the strong adsorption part from the extraction port C, supply the lyse d-II from the lyse supply port D-II, use the downstream end of the interval 4 as the extraction port B, extract the medium adsorption part from the extraction port B, use the upstream end of the interval 5 as the lyse supply port D -IV, supply the lyse d-IV from the lyse supply port D-IV, and extract the weak adsorption part from the extraction port A, thereby making the desorption force of the lyse flowing through the interval 1 the strongest, making the desorption force of the lyse flowing through the intervals 2, 3 and 4 weaker than the desorption force of the lyse flowing through the interval 1, and making the desorption force of the lyse flowing through the interval 5 weaker than the desorption force of the lyse flowing through the intervals 2, 3 and 4. A chromatographic separation system simulating a moving layer method is provided, which uses a circulation system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly through piping, and separates a weakly adsorbable component, a strongly adsorbable component, and a medium adsorbable component with an adsorption between the two components contained in a stock solution by using two or more solvents; the chromatographic separation system simulating a moving layer method is characterized in that: a stock solution supply port F, two ports corresponding to each of the two or more solvents are provided in the piping of the circulation system; The solvent supply port D on the bottom of the tower, the extraction port A for the weakly adsorbable part containing the weakly adsorbable component, the extraction port B for the medium adsorbable part containing the medium adsorbable component, and the extraction port C for the strong adsorbable part containing the strong adsorbable component; and the positions of the stock solution supply port F, the extraction port A, the extraction port B and the extraction port C are set as follows (a) to (c): (a) the extraction port B is set on the downstream side of the stock solution supply port F with at least one unit packed tower sandwiched therebetween; (b) the extraction port C is set at (c) the extraction port A is arranged in the piping having the extraction port B, or the extraction port A is arranged in the downstream side of the extraction port B, which is at least one unit packed tower; the stratification separation system is a simulated moving layer stratification separation system having a mechanism for sequentially repeating the following steps (A) and (B): [Step (A)] simultaneously or separately extracting the raw liquid from the raw liquid supply port F; The step of supplying a stock solution from the liquid supply port F, supplying two or more solvents from the two or more solvent supply ports D, and extracting a weakly adsorbable part from the extraction port A, a medium adsorbable part from the extraction port B, and a strongly adsorbable part from the extraction port C simultaneously or separately; [Step (B)] After the step (A) is completed, the stock solution supply port F, the solvent supply port D, the extraction port A, the extraction port B, and the extraction port C are moved downstream while maintaining their relative positional relationship.

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