TW202035004A - 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 moving layer chromatographic separation method and simulated moving layer chromatographic separation system

The invention relates to a chromatographic separation method in a simulated moving layer mode and a chromatographic separation system in a simulated moving layer mode.

In the chromatographic separation of the simulated moving layer method, a plurality of unit packed towers filled with adsorbents having selective adsorption capacity for a specific component of two or more components contained in the original solution (hereinafter also referred to as The "packed tower", sometimes called "pipe string"), is connected in series through piping, and the packed tower at the most downstream position is connected with the packed tower at the most upstream position to construct an endless circulation system. Supply the original solution and the eluent to the circulation system, and at the same time, the faster moving part (weakly adsorbent part) and the slower part (strongly adsorbent part) in the circulating system, and the medium moving part according to the need (Middle adsorptive part), respectively from different positions, and then make the original solution supply position, the eluent supply position, the extraction position of the weak adsorption part, the extraction position of the neutral adsorption part, and the extraction of the strong adsorption part Position, while maintaining a certain positional relationship, move to the fluid circulation direction of the circulatory system. By repeating this operation, the processing operation of the moving layer that can continuously supply the stock solution can be realized in a simulation. In Patent Document 1, it is disclosed that a series of improved simulated moving layer devices are used to repeatedly perform the steps of extracting the middle adsorptive part while supplying the eluent and the stock solution, and extracting the weakly adsorptive part and the strong while supplying the eluent. The step of the adsorptive part is a method of continuously separating 3 or more parts with different affinity to the adsorbent.

As represented by the technique described in Patent Document 1, basically one type of eluent is used in the conventional chromatographic separation of the general simulated moving layer method. Therefore, when supplying a stock solution containing components that are highly adsorbable to the adsorbent, or a stock solution containing components that are prone to tailing (a phenomenon in which the concentration distribution becomes wider) is supplied to the circulation system, in order to desorb these (Disengagement) Need to use a lot of eluent. The use of a large amount of eluent will increase the cost of concentration of the extract, and will also reduce the production volume of the adsorbent per unit of the target refined product.

On the other hand, in the chromatographic separation of the simulated moving layer method, two or more eluents are also used. For example, Patent Document 2 describes the use of a first eluent with a weak desorption force and a second eluent with a strong desorption force, and the timing of the supply of the eluent and the original solution and the weak adsorption The extraction timing of the part, the middle adsorption part, and the strong adsorption part is set to a specific combination, thereby achieving the technical content of higher separation efficiency with a smaller amount of adsorbent. [Prior Art] [literature] [Patent Literature]

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

[The problem to be solved by the invention]

The chromatographic separation of the simulating moving layer method can obtain the target refined object continuously and with high purity, so its application in medical and other fields is also reviewed by us. For example, in the manufacture of antibody drugs, in addition to the target antibody, in the extract or culture solution of cultured cells that produce antibodies, fragments that cannot fully function as antibodies due to the fragmentation of the antibody are also produced, or A large aggregate of antibodies. Generally speaking, the above-mentioned fragments have fewer sites for interaction with the adsorbent, and the adsorbability relative to the adsorbent is weak. On the contrary, the agglomerate has a strong adsorption property to the adsorbent. Therefore, when the chromatographic separation of the simulated moving layer method is applied to the purification of antibody drugs, the target antibody must be extracted as a mid-adsorptive part that shows moderate adsorptivity to the adsorbent. On the other hand, both the weakly adsorbing part and the strongly adsorbing part must be sufficiently removed at a high removal rate. In addition, in the practical process of these chromatographic separations, it is also very important to reduce the necessary amount of adsorbent as much as possible and improve the efficiency of separation treatment to achieve the purpose of cost reduction. However, the inventors of the present invention conducted a review of the conventional chromatographic separation using the simulated moving layer method, represented by the techniques described in the above-mentioned patent documents, and found that it is quite difficult to fully achieve the above-mentioned objective.

Therefore, the object of the present invention is to provide a chromatographic separation method using a simulated moving layer method, which can extract purification target components in a raw solution with a smaller amount of adsorbent with high purity. In addition, the object of the present invention is to provide a chromatographic separation system suitable for implementing the above-mentioned chromatographic separation method. [Means to solve the problem]

In view of the above-mentioned problems, the inventors of the present invention continue to focus on research and found that in the chromatographic separation method using the simulated moving layer method, two or more eluents are used, and the original solution in the circulation system is supplied to the port and The above-mentioned problems can be solved by setting the suction outlet for the adsorbent part, the suction outlet for the middle adsorbent part and the suction outlet for the weakly adsorbent part to a specific position. The present invention is based on such knowledge and has been continuously researched and completed.

The above-mentioned problems of the present invention are solved by the following means. 〔1〕 A method for chromatographic separation in a simulated moving layer method, which includes a circulation system in which a plurality of unit packed towers filled with adsorbents are connected in series and endlessly through pipes, and the raw liquid contained in the original solution is weaker than the adsorbent. The step of separating the adsorptive components, strong adsorptive components, and the adsorptive components in the middle of the two components with two or more eluents; the characteristic of this simulated moving layer chromatography separation method is: in the circulation system The piping is provided with a raw solution supply port F, two or more eluent supply ports D corresponding to each eluent of two or more types, a suction port A for the weakly adsorbable part containing the weakly adsorbable component, and The suction port B of the middle adsorptive portion of the middle adsorptive component and the suction port C of the strongly adsorptive portion containing the strong adsorptive component; the raw liquid supply port F, the suction port A, the suction port B, and the suction The position of the port C is set to the following (a) to (c): (a) The extraction port B is provided on the downstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (b) The pumping outlet C is provided in the piping having the raw liquid supply port F, or the pumping port C is provided on the upstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (c) the pumping outlet A, is installed in the piping with the extraction outlet B, or the extraction outlet A is installed on the downstream side of the extraction outlet B sandwiching at least one unit packed column; the chromatographic separation method includes sequential repeating The chromatographic separation method of simulated moving layer mode including the steps (A) and (B): [Step (A)] The stock solution is supplied from the stock solution supply port F at the same time or separately, two or more types of eluent are supplied from the two or more eluent supply ports D, and the weak adsorption is drawn from the extraction port A at the same time or separately. Part, the step of extracting the medium adsorptive part from the extraction port B and extracting the strong adsorptive part from the extraction port C; [Step (B)] After the step (A) is completed, the raw liquid supply port F, the eluent supply port D, the pumping port A, the pumping port B, and the pumping port C are made to move downstream while maintaining their relative positional relationship Steps to move sideways. 〔2〕 [1] The simulated moving layer chromatography 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 a stock solution; and a sub-step of not supplying the stock solution . [3] As described in [1] or [2], the chromatographic separation method of the simulated moving layer method, wherein the extraction port C is set to supply the eluent d1 with the strongest desorption power among the two or more eluents. On the downstream side of the eluent supply port D1; from the eluent supply port D1 to the extraction port C at least one unit packed tower is arranged; in this step (A), the eluent d1 is supplied During the period of time, from the suction port C, the same amount of the strongly adsorbed portion as the supply amount of the eluent d1 is extracted. [4] As described in any one of [1] to [3], the chromatographic separation method of the simulated moving layer method, wherein the extraction port B is provided at the second desorption power in supplying two or more eluents On the downstream side of the eluent supply port D2 of the strong eluent d2; at least one unit packed tower is arranged from the eluent supply port D2 to the suction port B; in this step (A), set During the supply of the eluent d2, a time zone of the middle adsorptive portion of the same amount as the supply amount of the eluent d2 is extracted from the suction port B. [5] The pseudo-moving-layer chromatographic separation method described in any one of [1] to [4], wherein 4 to 6 types of eluents with different desorption powers are used. [6] As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 4 or more unit packed towers, and the circulation system has at least 1 in each section The unit packed tower is divided into 4 sections 1 to 4 connected in an annular shape from the upstream side to the downstream side. In addition, two or more types of eluents are used, and the following is performed in this step (A) Sub-steps (A1-1), (A2-1) and (A3-1): ＜Substep (A1-1)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, and uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of section 3 The end is used as the raw liquid supply port F, the raw liquid is supplied from the raw liquid supply port F, the downstream end of the section 4 is used as the suction port A, and the weakly adsorbing part is drawn from the suction port A, thereby allowing the flow of the section 1 The desorption force of the eluent is the strongest. The desorption force of the eluent flowing in zone 2 is weaker than that of the eluent flowing in zone 1, so that the desorption of the eluent flowing in zones 3 and 4 is weaker. The desorption force is weaker than the desorption force of the eluent flowing in zone 2; ＜Sub-step (A2-1)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The upstream end of the section 3 serves as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the suction port A, thereby making the section The desorption force of the eluent flowing in 1 is the strongest. The desorption force of the eluent flowing in section 2 is weaker than that of the eluent flowing in section 1. The desorption force of the eluent is weaker than the desorption force of the eluent circulating in zone 2; <Sub-step (A3-1)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 3 is used as the suction port B, and the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 4 is used as the eluent supply port D-IV, from the eluent supply port D-IV The eluent solution d-IV is supplied, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and the eluent flowing in sections 2 and 3 are desorbed. The adhesion force is weaker than the desorption force of the eluent flowing in section 1, and the desorption force of the eluent flowing in section 4 is weaker than that of the eluent flowing in sections 2 and 3. [7] As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 4 or more unit packed towers, and the circulation system has at least 1 in each section The unit packed tower is divided into 4 sections 1 to 4 connected in an annular shape from the upstream side to the downstream side. In addition, two or more types of eluents are used, and the following is performed in this step (A) Sub-steps (A1-2), (A2-2) and (A3-2): ＜Substep (A1-2)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The raw liquid supply port F supplies raw liquid, and the downstream end of the section 4 is used as the suction port A, and the weakly adsorptive part is drawn from the suction port A, thereby increasing the desorption power of the eluent flowing in the sections 1 and 2 The strongest, making the desorption force of the eluent flowing in sections 3 and 4 weaker than the desorption force of the eluent flowing in sections 1 and 2; <Sub-step (A2-2)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, and uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of section 3 The end is used as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the eluent flowing in the zone 1 The desorption power of the eluent is the strongest. The desorption force of the eluent flowing in section 2 is weaker than that of the eluent flowing in section 1, and the desorption of the eluent flowing in sections 3 and 4 The adhesion force is weaker than the desorption force of the eluent flowing in zone 2; <Substep (A3-2)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-2) For the eluent d-II, the downstream end of section 3 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of section 4 is used as the eluent supply port D-IV from The eluent supply port D-IV supplies the eluent d-IV, and the weakly adsorptive part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in the section 1 is maximized, and the sections 2 and 3 The desorption force of the eluent flowing in zone 3 is weaker than that of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 4 is higher than that of zones 2 and 3. The desorption force of the liquid is weaker. 〔8〕 As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 5 or more unit packed towers, and the circulation system has at least 1 in each section The unit packing tower is divided into five sections 1 to 5 connected in a ring shape from the upstream side to the downstream side. In addition, the two or more types of eluents are used, and the following steps are performed in this step (A) Steps (A1－3), (A2－3) and (A3－3): <Sub-step (A1-3)> The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 serves as the eluent supply port D-III, the eluent supply port D-III supplies eluent d-III, and the downstream end of section 5 As the extraction port A, the weakly adsorbing part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in sections 1 and 2 is maximized, and the desorption force of the eluent flowing in section 3 The desorption power of the eluent flowing in zones 1 and 2 is the same or weaker than that of the eluent flowing in zones 1 and 2, so that the desorption power of the eluent flowing in zones 4 and 5 is The desorption force is weaker than that of the dissolving liquid circulating in zone 3; ＜Substep (A2－3)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II, and supplies it from the eluent Port D-III supplies the eluent d-III, and extracts the weakly adsorbing part from the extraction port A, thereby maximizing the desorption force of the eluent flowing in section 1 and making the leaching liquid circulated in sections 2 and 3 strongest The desorption force of the eluent is weaker than the desorption force of the eluent flowing in zone 1, and the desorption force of the eluent flowing in zones 4 and 5 is higher than that of the eluent flowing in zones 2 and 3. Weaker desorption; ＜Substep (A3－3)＞ The eluent d-I is supplied from the eluent supply port D-1, the strongly adsorbing part is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-3) For the eluent d-II, the downstream end of the section 4 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV. The eluent supply port D-IV supplies the eluent d-IV, and the weakly adsorptive part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and section 2, The desorption force of the eluent flowing in 3 and 4 is weaker than the desorption force of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 5 is higher than that of zones 2, 3 and 4. The desorption force of the circulating solution is weaker. 〔9〕 As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 7 or more unit packed towers, and the circulation system has at least 1 in each section The unit packing tower is divided into five sections 1 to 5 connected in a ring shape from the upstream side to the downstream side. In addition, using these two or more eluents, perform the following in this step (A) Sub-steps (A1-4), (A2-4) and (A3-4): ＜Substep (A1-4)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 serves as the eluent supply port D-III, the eluent supply port D-III supplies eluent d-III, and the downstream end of section 5 As the extraction port A, the weakly adsorbing part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in sections 1 and 2 is maximized, and the desorption force of the eluent flowing in section 3 The desorption power of the eluent flowing in zones 1 and 2 is the same or weaker than that of the eluent flowing in zones 1 and 2, so that the desorption power of the eluent flowing in zones 4 and 5 is The desorption force is weaker than that of the dissolving liquid circulating in zone 3; ＜Substep (A2－4)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of the section 4 The end is used as the eluent supply port D-IV, the eluent d-IV is supplied from the eluent supply port D-IV, and the weakly adsorptive part is extracted from the extraction port A, thereby making the eluent flowing in the zone 1 The desorption force of the eluent is the strongest, making the desorption force of the eluent flowing in sections 2 and 3 weaker than the desorption force of the eluent flowing in the section 1, making the eluent flowing in the sections 4 and 5 weaker The desorption force of is weaker than the desorption force of the dissolving liquid circulating in intervals 2 and 3; <Substep (A3-4)> The eluent d-I is supplied from the eluent supply port D-1, the strongly adsorbing part is extracted from the extraction port C, and supplied from the eluent supply port D-II in the substep (A2-4) For the eluent d-II, the downstream end of the section 4 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-V, from The eluent supply port D-V supplies the eluent d-V, and the weakly adsorbing part is extracted from the extraction port A, thereby making the desorption force of the eluent flowing in section 1 strongest, making section 2. The desorption force of the eluent flowing in 3 and 4 is weaker than the desorption force of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 5 is higher than that of zones 2, 3 and 4. The desorption force of the circulating solution is weaker. [10] As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 5 or more unit packed towers, and the circulation system has at least 1 in each section The unit packing tower is divided into five sections 1 to 5 connected in a ring shape from the upstream side to the downstream side. In addition, using these two or more eluents, perform the following in this step (A) Sub-steps (A1-5), (A2-5) and (A3-5): <Substep (A1-5)> The upstream end of the section 3 is used as the stock solution supply port F, the stock solution is supplied from the stock solution supply port F, and the upstream end of the section 4 is used as the eluent supply port D-III, from the eluent supply port D-III The eluent solution d-III is supplied, and the downstream end of the section 5 is used as the suction port A, and the weakly adsorbable part is extracted from the suction port A, whereby the desorption force of the eluent liquid flowing in the section 3 is maximized. Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; ＜Substep (A2－5)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II, and supplies it from the eluent Port D-III supplies the eluent d-III, and extracts the weakly adsorbing part from the extraction port A, thereby maximizing the desorption force of the eluent flowing in section 1 and making the leaching liquid circulated in sections 2 and 3 strongest The desorption force of the eluent is weaker than the desorption force of the eluent flowing in zone 1, and the desorption force of the eluent flowing in zones 4 and 5 is higher than that of the eluent flowing in zones 2 and 3. Weaker desorption; ＜Substep (A3－5)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 4 serves as the suction port B, and the middle adsorptive part is extracted from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV, and from the eluent supply port D-IV The eluent d-IV is supplied, and the weakly adsorbing part is extracted from the suction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and the eluent flowing in sections 2, 3 and 4 The desorption power of is weaker than that of the eluent flowing in zone 1, and the desorption power of the eluent flowing in zone 5 is higher than that of the eluent flowing in zones 2, 3 and 4. weaker. [11] As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 5 or more unit packed towers, and the circulation system has at least 1 in each section The unit packing tower is divided into five sections 1 to 5 connected in a ring shape from the upstream side to the downstream side. In addition, using these two or more eluents, perform the following in this step (A) Sub-steps (A1-6), (A2-6) and (A3-6): ＜Substep (A1－6)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the downstream end of section 3 is used as the suction port B. The suction port B draws out the middle adsorptive part, and uses the upstream end of section 4 as the eluent supply port D-IV, and supplies the eluent d-IV from the eluent supply port D-IV to the downstream side of section 5 The end is used as the extraction port A, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in sections 1, 2 and 3 is maximized, and the eluent flowing in sections 4 and 5 The desorption force of the liquid is weaker than the desorption force of the dissolving liquid circulating in zones 1, 2 and 3; <Substep (A2-6)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 3 as the stock solution supply port F, supplies the stock solution from the stock solution supply port F, and uses the upstream end of the section 4 as the eluent supply port D-III, from The eluent supply port D-III supplies the eluent d-III, and the weakly adsorbable part is extracted from the extraction port A, thereby making the desorption force of the eluent flowing in section 1 the strongest, making the section 3 The desorption force of the circulating eluent is weaker than the desorption force of the eluent circulating in section 1, so that the desorption force of the eluent circulating in sections 4 and 5 is higher than that of the eluent circulating in section 3. Weaker desorption; <Substep (A3-6)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the interval 1 The desorption force of the eluent flowing in 1 is the strongest, and the desorption force of the eluent flowing in sections 2 and 3 is weaker than that of the eluent flowing in section 1. The desorption force of the circulating eluate is weaker than the desorption force of the circulating eluate in sections 2 and 3. [12] As described in any one of [1] to [5], the chromatographic separation method of the simulated moving layer method, wherein the circulation system has 5 or more unit packed towers, and the circulation system has at least 1 in each section The unit packing tower is divided into five sections 1 to 5 connected in a ring shape from the upstream side to the downstream side. In addition, using these two or more eluents, perform the following in this step (A) Sub-steps (A1-7), (A2-7) and (A3-7): ＜Substep (A1-7)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 3 as the stock solution supply port F, supplies the stock solution from the stock solution supply port F, and uses the upstream end of the section 4 as the eluent supply port D-III, from The eluent supply port D-III supplies the eluent d-III, and the downstream end of the section 5 is used as the suction port A, and the weakly adsorbing part is extracted from the suction port A, thereby making the flow in the section 1 The desorption force of the eluent is the strongest. The desorption force of the eluent flowing in zone 3 is weaker than that of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zones 4 and 5 is weaker. The desorption force is weaker than the desorption force of the eluent circulating in zone 3; <Substep (A2-7)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the interval 1 The desorption force of the eluent flowing in 1 is the strongest, and the desorption force of the eluent flowing in sections 2 and 3 is weaker than that of the eluent flowing in section 1. The desorption force of the circulating eluent is weaker than the desorption force of the circulating eluent in sections 2 and 3; <Substep (A3-7)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 4 serves as the suction port B, and the middle adsorptive part is extracted from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV, and from the eluent supply port D-IV The eluent d-IV is supplied, and the weakly adsorbing part is extracted from the suction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and the eluent flowing in sections 2, 3 and 4 The desorption power of is weaker than that of the eluent flowing in zone 1, and the desorption power of the eluent flowing in zone 5 is higher than that of the eluent flowing in zones 2, 3 and 4. weaker. [13] A simulated moving-layer chromatography separation system, which uses a plurality of unit packed towers filled with adsorbents in a circulatory system connected in series and endlessly through piping to weakly adsorb the raw liquid contained in the adsorbent against the adsorbent The chromatographic separation system of the simulated moving layer method is characterized by: the piping in the circulation system, which is separated by two or more eluents. , Provided with the original solution supply port F, two or more eluent supply ports D corresponding to each eluent of two or more kinds, the suction port A of the weakly adsorbable part containing the weakly adsorbable component, and the medium adsorbing The suction port B of the mid-adsorptive portion of the sexual component and the suction port C of the strong-absorptive portion containing the strong adsorptive component; the raw liquid supply port F, the pumping port A, the pumping port B, and the pumping port C The position is set as the following (a) ~ (c): (A) The extraction port B is provided on the downstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (B) The pumping port C is provided in the piping having the raw liquid supply port F, or the pumping port C is provided on the upstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (c) ) The extraction outlet A is provided in the piping with the extraction outlet B, or the extraction outlet A is provided on the downstream side of the extraction outlet B sandwiching at least one unit packed tower; the chromatographic separation system A simulated moving layer chromatography separation system with a mechanism that repeats the following steps (A) and (B) in sequence: [Step (A)] The stock solution is supplied from the stock solution supply port F at the same time or separately, two or more types of eluent are supplied from the two or more eluent supply ports D, and the weak adsorption is drawn from the extraction port A simultaneously or separately Part, the step of extracting the medium adsorptive part from the extraction port B and extracting the strong adsorptive part from the extraction port C [Step (B)] After the step (A) is completed, the raw liquid supply port F, the eluent supply port D, the pumping port A, the pumping port B, and the pumping port C are made to move downstream while maintaining their relative positional relationship Steps to move sideways.

In this manual, the terms "upstream" and "downstream" apply to the flow direction of the fluid in the circulation system. That is, for a certain part of the circulatory system, the so-called "upstream side" refers to the side from which fluid flows to that part, and the so-called "downstream side" refers to the side from which fluid flows out. In this specification, the so-called "strongly adsorptive component" refers to the component that has a strong adsorption force with respect to the adsorbent among the plural components contained in the original liquid; Among the plural components contained in the sorbent, the component that has a weaker adsorption force with respect to the adsorbent; the so-called "medium adsorptive component" means that the adsorptive property to the adsorbent is weaker than the above-mentioned strong adsorptive component and is relatively A component that has stronger adsorptivity than the aforementioned weakly adsorptive component. That is, the terms "strong adsorption", "medium adsorption" and "weak adsorption" show the relative strength of the adsorption force when comparing the adsorption force of each component contained in the original solution with respect to the adsorbent. The above-mentioned "strongly adsorptive components", "medium adsorptive components" and "weakly adsorptive components" may each be composed of a single component or may be composed of a plurality of components. In addition, the adsorption power of the plural components may be the same or different. The grouping of "strongly adsorbable components", "medium adsorbent components" and "weakly adsorbable components" of each component in the original solution can be set appropriately according to the purpose. Taking the situation where the original solution contains 4 components as an example, the two components that are ranked stronger with respect to the adsorbent can be assigned the position of strong adsorbent together, which is the third strongest for the adsorbent. The composition of the sorbent is given the status of the middle adsorptive component, and the component with the weakest adsorption force relative to the adsorbent is given the status of the weak adsorptive component. In addition, the component with the strongest adsorption force to the adsorbent can also be given the position of strong adsorbent component, and the second strongest component with respect to the adsorbent and the third strongest component can be given the middle adsorbability. The status of the component, and the weakest adsorption component is assigned to the component with the weakest adsorption force relative to the adsorbent. In addition, the component with the strongest adsorption force relative to the adsorbent can also be assigned the position of the strong adsorbent component, and the component with the second strongest adsorption force relative to the adsorbent can be assigned the position of the middle adsorbent component. The third strongest component and the weakest component of the adsorbent are given the position of the weakest component. The original solution contains more than 5 kinds of components, and it can also be separated and refined based on various groups. In the present invention, the so-called "desorption force" of the eluent is the strength of the action of instructing the components adsorbed by the adsorbent to desorb from the adsorbent. [Effect of Invention]

According to the chromatographic separation method of the simulated moving layer method of the present invention, it is possible to extract the purification target components in the stock solution with high purity while suppressing the amount of adsorbent used. In addition, the simulated moving layer chromatographic separation system of the present invention can be suitably used in the implementation of the simulated moving layer chromatographic separation method of the present invention.

Hereinafter, a preferred embodiment of the chromatographic separation method of the simulated moving layer method of the present invention (hereinafter also referred to as "the method of the present invention") is described.

The method of the present invention is implemented using a circulation system in which a plurality of unit packed towers filled with adsorbents are connected in series and endlessly through piping. The circulation system body used in the simulated moving layer method is well-known to the public. For example, refer to Japanese Patent Publication No. 2009-36536 or Japanese Patent No. 4606092. Hereinafter, the circulatory system is illustrated with a diagram, but the present invention is not limited to these implementation aspects except for the content limited by the present invention. In addition, the drawings disclosed below are explanatory diagrams to make the present invention easier to understand. The size of each structure or the relative size relationship may change for convenience of explanation, and it does not show the actual relationship as it is. In addition, it is not limited to the shape, relative positional relationship, etc. disclosed in the drawings except for the matters limited by the present invention. In addition, conditions other than those defined by the present invention, such as the capacity of the unit packed tower, the internal cross-sectional area or length of the pipe, and the flow rate of the liquid supplied to the circulation system, 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 FIG. 1. The circulation system 100 shown in Fig. 1 is equipped with 4 unit packed towers (tube columns) filled with adsorbent Ab (unit packed towers 10a, 10b, 10c, 10d), and the outlet of each unit packed tower is connected to At the entrance of adjacent unit packing towers, each unit packing tower is connected in series to form a whole. Then, the outlet of the last unit packed tower (for example, the unit packed tower 10d) is connected to the inlet of the foremost unit packed tower (for example, the unit packed tower 10a) through a pipe 111, and all the unit packed towers are connected in an endless shape (ring shape). With these structures, the fluid can be circulated in the circulation system 100. The unit packed towers 10a to 10d may have the same shape, size, and filling amount of the adsorbent, or they may be different. For the unit packed tower 10a-10d, the internal shape, size, and filling amount of the adsorbent should all be equivalent (more preferably the same).

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

Between the shutoff valves R1 to R4 and the outlets of the unit packed towers 10a to 10d located on the upstream side of the shutoff valves R1 to R4 are respectively arranged in a branched manner that will contain a large amount of weakly adsorbent components relative to the adsorbent Ab. Part (in this specification, the "weakly adsorbable part relative to the adsorbent Ab" is also simply referred to as the "weakly adsorbable part") the extracted weakly adsorbent part is drawn out of the lines 2a, 2b, 2c, 2d. The weakly adsorbing part extraction lines 2a, 2b, 2c, 2d are respectively provided with weakly adsorbing part extraction valves A1, A2, A3, A4 that can open and close the weakly adsorbing part extraction lines. The weakly adsorptive parts are drawn out of the pipelines 2a, 2b, 2c, and 2d, merge and merge into a weakly adsorptive part confluence pipe 2J.

In addition, similarly, between the shutoff valves R1 to R4 and the outlets of the unit packed towers 10a to 10d located on the upstream side thereof, there are arranged in a branched manner to contain a large amount of the adsorbent Ab. The part of the adsorptive component (in this specification, the "neutral adsorptive part with respect to the adsorbent Ab" is also simply referred to as the "intermediate adsorptive part"), the middle adsorptive part is extracted from the lines 3a, 3b, 3c, 3d. The middle adsorptive part extraction lines 3a, 3b, 3c, 3d are respectively provided with middle adsorptive part extraction valves B1, B2, B3, B4 that can open and close each of the middle adsorptive part extraction lines. The middle adsorptive parts are drawn out of the pipelines 3a, 3b, 3c, and 3d, merged and collected into a middle adsorptive part confluence pipe 3J.

In addition, similarly, between the shutoff valves R1 to R4 and the outlets of the unit packed towers 10a to 10d located on the upstream side of the shutoff valves R1 to R4 and the outlets of the unit packing towers 10a to 10d located on the upstream side thereof, there are arranged in a branched manner to contain a large amount of strong The part of the adsorbent component (the "strongly adsorbent part relative to the adsorbent Ab" is also simply referred to as the "strongly adsorbent part" in this specification) is extracted from the lines 4a, 4b, 4c, 4d. The strong adsorption part extraction lines 4a, 4b, 4c, 4d are respectively provided with strong adsorption part extraction valves C1, C2, C3, C4 that can open and close the strong adsorption part extraction lines. Each strong adsorption part is drawn out of the pipelines 4a, 4b, 4c, 4d, merged and gathered into a strong adsorption part junction pipe 4J.

In the step (A) described later, any one of the weakly adsorbing partial extraction valves A1, A2, A3, and A4 is in an open state. The connection part of the weakly adsorbent part extraction line and the piping 111 provided with this open weakly adsorbent part extraction valve is the weakly adsorbent part extraction port A in the step (A) described later. In addition, in the step (A) described later, any one of the middle adsorptive partial extraction valves B1, B2, B3, and B4 is in an open state. The connection part of the middle adsorptive part extraction line and the piping 111 where the adsorptive part extraction valve in the open valve is provided is the middle adsorptive part extraction port B in the step (A) described later. In addition, in the step (A) described later, any one of the highly adsorptive partial extraction valves C1, C2, C3, and C4 is in an open state. The connection part of the strong adsorption part extraction line and the piping 111 provided with this valve-opening strong adsorption part extraction valve is the strong adsorption part extraction port C in the step (A) mentioned later.

In the circulatory system 100, in order to prevent the pressure of the circulatory system 100 from increasing excessively, a safety valve (or a pressure reducing valve) not shown in the figure may be installed at an appropriate position. In addition, it is advisable to install check valves T1, T2, T3, and T4 to prevent backflow between two adjacent unit packed towers.

In the circulation system 100, as shown in FIG. 1, a structure capable of supplying the stock solution 117 stored in the stock solution tank 116 is configured. In addition, the circulation system 100 has a structure capable of supplying two or more types of eluent. As an example, Fig. 1 shows a state where four types of eluent are supplied. The stock solution 117 is supplied through the stock solution supply line 11 by the stock solution supply pump P2 that can control the supply flow rate. The original liquid supply pump P2 should be a quantitative pump. The stock solution supply line 11, as shown in Fig. 1, constitutes "four stock solution supply branch lines 11a, 11b, 11c, and 11d branched out, through each stock solution supply branch line 11a, 11b, 11c, 11d, the stock solution can be separately supplied to Each unit packing tower 10a, 10b, 10c, 10d entrance" structure. Each of the original solution supply branch lines 11a, 11b, 11c, 11d is provided with openable and closable original solution supply valves F1, F2, F3, F4, and the original solution is supplied to the downstream through the original solution supply branch line with the open valve of the original solution supply valve The connected unit fills the tower. In step (A) described later, any one of the above-mentioned stock solution supply valves F1, F2, F3, and F4 is in an open state. The connection part of the original solution supply branch line and the piping 111 provided with the open solution supply valve is the original solution supply port F in the step (A) described later.

Figure 1 shows the state of supplying four eluents with different desorption powers. The eluent 9a stored in the eluent tank 8a is supplied to the eluent supply line 12 by the eluent supply pump P3 whose supply flow rate can be controlled. The eluent 9b stored in the eluent tank 8b is supplied to the eluent supply line 13 by the eluent supply pump P4 whose supply flow rate can be controlled. The eluent 9c stored in the eluent tank 8c is supplied to the eluent supply line 14 by the eluent supply pump P5 whose supply flow rate can be controlled. Furthermore, the eluent 9d stored in the eluent tank 8d is supplied to the eluent supply line 15 by the eluent supply pump P6 whose supply flow rate can be controlled. The eluent supply pumps P3 to P6 should preferably be quantitative pumps. The eluent supply line 12, as shown in FIG. 1, constitutes "four eluent supply branch lines 12a, 12b, 12c, 12d, and each eluent supply branch line 12a, 12b, 12c, 12d is branched, The eluate can be supplied to the inlet of each unit packed tower 10a, 10b, 10c, and 10d." Each eluent supply branch line 12a, 12b, 12c, 12d is provided with openable and closable eluent supply valves E1a, E2a, E3a, E4a, and the eluent supply branch is provided with an open valve. The pipeline supplies the eluent to the unit packed tower connected downstream. Similarly, the eluent supply line 13 branches into four eluent supply branch lines 13a, 13b, 13c, and 13d, and the eluent supply line 14 branches into four eluent supply branch lines 14a, 14b, 14c, 14d. , The eluent supply line 15 branches into four eluent supply branch lines 15a, 15b, 15c, 15d, and constitutes a structure that can supply each eluent to the inlet of each unit packed tower 10a, 10b, 10c, 10d . The eluent supply branch lines 13a, 13b, 13c, 13d are provided with openable and closable eluent supply valves E1b, E2b, E3b, E4b, and the eluent supply branch lines 14a, 14b, 14c, 14d, respectively The eluent supply valves E1c, E2c, E3c, and E4c that can be opened and closed are provided, and the eluent supply valves E1d, E2d, E3d, E3d, E3d, E3d, E3d, E3d, E3d and E4d. In the step (A) described later, the connection portion of the eluent supply branch line with the open eluent supply valve and the pipe 111 is the eluent supply port D. In the method of the present invention, two or more types of eluents are used, so in the step (A) described later, the eluent supply valves to be opened are plural. Therefore, in the step (A) described later, the eluent supply port D has a number (two or more) corresponding to the type of eluent used.

Next, the operation of the circulatory system when the method of the present invention is implemented using the above circulatory system will be described. However, the present invention is not limited to these implementation aspects except for the content limited by the present invention. In the method of the present invention, the circulation system is such that the positions of the raw liquid supply port F, the weakly adsorbing part extraction port A, the medium adsorbent part pumping outlet B, and the strong adsorbent part pumping outlet C are formed to satisfy the following (a)~ (C) Relationship. That is, in the repetition of steps (A) and (B) described later, the relative positional relationship between the original liquid supply port F, the weakly adsorbent part extraction port A, the middle adsorbent part extraction port B, and the strong adsorbent part extraction port C , Often meet the (a) ~ (c).

(A) The middle adsorptive part extraction port B is provided on the downstream side of the raw liquid supply port F sandwiching at least one unit packing tower.

(B) Install the strong adsorption part extraction port C in the piping with the raw liquid supply port F, or set the strong adsorption part extraction port C at the upstream of the raw liquid supply port F sandwiching at least one unit packing tower side. Here, the phrase "install the strong adsorption part extraction port C in the piping with the stock solution supply port F" means that there is no unit packing tower arranged between the strong adsorption part extraction port C and the stock solution supply port F meaning. In addition, in the case of "installing the highly-absorptive portion extraction port C in the pipe having the undiluted solution supply port F", the strongly-absorbent portion evacuating port C is provided on the upstream side of the same piping compared to the original solution supply port F. This corresponds to the overall relationship between the extraction port and the supply port provided in the same pipe. That is, in a circulation system, when a certain extraction port and a supply port are installed in the same piping (when the extraction port and the supply port are not sandwiched between the unit packing tower), compared with the supply port, the extraction The ports are arranged on the upstream side of the same pipe. This is to prevent the supplied liquid from being pumped out from the pumping outlet before it reaches the downstream unit packed tower. The above-mentioned (b) is preferably an aspect in which "exhaust port C of the strong adsorbent portion is provided on the upstream side of the raw liquid supply port F sandwiching at least one unit packed tower".

(C) Install the weak adsorption part extraction port A in the piping with the middle adsorption part extraction port B, or set the weak adsorption part extraction port A to at least 1 Pack the downstream side of the tower. Here, the so-called "install the weakly adsorbent part extraction port A in the pipe with the middle adsorbent part extraction port B" means between the weakly adsorbent part extraction port A and the middle adsorbent part extraction port B, and It means that there is no unit packed tower. The above-mentioned (c) is preferably an aspect in which "weakly adsorbent part extraction port A is provided on the downstream side of middle adsorbent part extraction port B sandwiching at least one unit packed tower".

In the method of the present invention, the following steps (A) and (B) are sequentially repeated using the above-mentioned circulation system.

[Step (A)] Supply the stock solution from the stock solution supply port F at the same time or separately, and supply more than two types of eluent from the two or more eluent supply ports D, and draw out the weak adsorption from the weakly adsorbing part extraction port A at the same time or separately The step of extracting the middle adsorptive part from the middle adsorptive part extraction port B, and extracting the strong adsorptive part from the strong adsorptive part extraction port C. 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 eluent solution supplied from the eluent supply port D, and the amount of the eluate from the weakly adsorbing portion A The total amount of the extracted weakly adsorptive portion, the amount of the intermediate adsorptive portion extracted from the outlet B of the middle adsorptive portion, and the amount of the strong adsorptive portion C extracted from the outlet C of the strongly adsorptive portion are the same. That is, in a state where the liquid is supplied into the circulation system, the same amount of liquid is drawn from the circulation system. To explain in more detail, when liquid is supplied from a certain supply port (X) and liquid is drawn from a certain outlet (Y) on the downstream side, if it is not from the downstream side of X and the upstream side of Y Where liquid is supplied, the amount of liquid drawn from Y is the same as the amount of liquid supplied from X. In addition, if the liquid is supplied from a place downstream from X and upstream from Y, the amount of liquid drawn from Y and the amount of liquid supplied from X are the same as those from the downstream side of X and more than Y The total amount of the liquid supplied to the further upstream side is the same. For example, in the sub-step (A1-1) of Fig. 2, the supply amount of the eluent supplied from the eluent supply port D1, and the extraction of the strongly adsorbent part extracted from the strongly adsorbent part extraction port C The same amount. In addition, in the same substep (A1-1), the sum of the supply amount of the eluent supplied from the eluent supply port D2 and the supply amount of the original liquid supplied from the original liquid supply port F, and the weak adsorption The amount of the weakly adsorbing part extracted from the partial extraction port A is the same. In addition, the above-mentioned "simultaneous or separate supply" means supplying with no time difference (not staggering the supply timing) or supplying with a timing difference (staggering the supply timing). Among them, in one step (A), when two or more supply ports for supplying two or more liquids are arranged in the same piping (when the two or more supply ports are arranged in a manner that does not sandwich the unit packed tower When piping and supplying different liquids from the two or more supply ports), the supply of the two or more liquids will not be performed at the same time. That is, in one step (A), the supply of the two or more liquids is performed in different substeps. Similarly, in one step (A), when two or more extraction ports for extracting two or more types of parts are arranged in the same piping (when the two or more extraction ports do not sandwich the unit packed tower When it is arranged in the piping and different parts are extracted from each of the two or more extraction ports), the extraction of the two or more types of parts will not be performed at the same time. That is, in one step (A), the extraction of the two or more types of parts is performed in different substeps.

[Step (B)] After the end of this step (A), make the original solution supply port F, the eluent supply port D, the weakly adsorbent portion pumping port A, the medium adsorbent portion pumping port B, and the strong adsorbent portion pumping port C, while maintaining their relative Steps to move to the downstream side in a positional relationship. This movement to the downstream side is to instruct the raw solution supply port F, the eluent supply port D, the weakly adsorbent part pumping port A, the medium adsorbent part pumping port B, and the strong adsorbing part pumping port C to maintain their relative positions. In the state of the relationship, it means to move the unit to the downstream side and fill the tower by 1 unit. For example, in step (A), when the connecting portion of the original solution supply branch line with the original solution supply valve F1 and the pipe 111 is the original solution supply port F, the original solution supply port F is moved to The connection part of the original solution supply branch line and the pipe 111 where the original solution supply valve F2 is provided. This is the same for the eluent supply port D, the weakly adsorbent part extraction port A, the middle adsorbent part pumping port B, and the strong adsorbent part pumping port C. In addition, the movement of one unit of the unit packed tower on the downstream side of the supply ports and the suction ports described above can be performed by adjusting the opening and closing of various pumps or valves arranged in the circulation system. Through the implementation of step (B), in the next step (A) [referred to as step (A2)], and the step (A) before this step (B) [referred to as step (A1)] In the same way as for each unit packed tower, the liquid is supplied and extracted, and the comparison step (A1) is carried out to each unit packed tower one unit downstream.

The above step (A) should be composed of multiple sub-steps. At this time, the supply of the original solution from the original solution supply port F, the supply of two or more types of eluents from the two or more eluent supply ports D, and the extraction of the weakly adsorbed portion of the port A from the weakly adsorbed portion Which sub-step is used for extraction, extraction of the intermediate adsorption part from the outlet B of the middle adsorption part, and extraction of the strong adsorption part from the exit C of the strong adsorption part, can be carried out within the scope of not impairing the efficacy of the present invention Within, set appropriately according to the purpose. In particular, in the method of the present invention, the above-mentioned step (A) preferably includes the sub-step of supplying the stock solution and the sub-step of not supplying the stock solution. That is, in step (A), it is preferable that there is a time when the stock solution is supplied and a time when the stock solution is not supplied.

In step (A), two or more eluents are supplied. It is preferable to provide the supply port of the eluent with the strongest desorption force among the two or more types of eluents in the piping on the upstream side of the unit packed tower with respect to the extraction port C of the strong adsorptive portion. It is advisable to install the remaining eluent supply port and even the stock solution supply port in the same pipe as the supply port of the eluent with the strongest desorption force, or install it compared to the supply port of the eluent with the strongest desorption force. On the more upstream side sandwiching the unit packing tower. The so-called "installed in the same piping as the supply port of the eluent with the strongest desorption force, or installed on the upstream side sandwiching the unit packed tower compared to the supply port of the eluent with the strongest desorption force." Refers to the piping with the strongest eluent supply port as the starting point and proceed to the upstream side until it reaches the piping with the strong absorption part extraction port C. The strongest eluent is installed in any of the piping in between. The meaning of the supply port of the eluent other than the liquid or even the supply port of the raw liquid (in other words, it refers to the downstream side from the piping provided with the suction port C of the strong adsorptive part as the starting point, until it reaches the supply of the strongest eluent It means that the strongest eluent other than the eluent supply port or even the original solution supply port is provided in any one of the piping in between. To put it another way, it means that when the strongest eluent is used When two or more unit packed towers are installed between the supply port of and the suction port C of the strong adsorptive part, the piping that connects the two or more unit packed towers does not provide the eluent supply port or the raw liquid supply port the meaning of). At this time, if there are two or more types of eluents other than the strongest eluent, two or more supply ports corresponding to these eluents can be installed in the same piping, or each can be installed in the sandwich unit. Individual piping for the tower. In addition, the supply port of the eluent other than the strongest eluent and the supply port of the stock solution may be provided in the same pipe. In addition, it is preferable to install one of the supply ports for the eluent liquid other than the eluent with the strongest desorption force in the same pipe as the supply port for the eluent with the strongest desorption force. An implementation aspect.

In step (A), it is appropriate to "drain the supply amount of the eluent with the strongest desorption force from the downstream during the supply of the eluent with the strongest desorption force among the two or more eluents. The same amount of strong adsorption part". At this time, it is advisable to arrange at least one unit packed tower from the supply port of the eluent with the strongest desorption force to the suction port of the strongly adsorbing part downstream of it, and there is no other supply port in between. The appearance.

In step (A), it is advisable to supply the eluent with the second strongest desorption force among the two or more eluents, and to draw out the eluent with the second strongest desorption force from its downstream. The time zone of the adsorptive part of the same amount of supply (substep). At this time, at least one unit packed tower (preferably a plurality of unit packed towers) is arranged from the supply port of the second strongest eluent with the second strongest desorption force to the outlet of the middle adsorptive part downstream of it. In addition, even if there are other supply ports between the piping provided with the supply port for the eluent with the second strongest desorption force and the piping provided with the suction port for the middle adsorptive portion on the downstream side, the above-mentioned The liquid is supplied from the other supply port during the time period during which the absorbent part is extracted.

During the implementation of step (A), it is advisable to often extract the weakly adsorbing part. Therefore, when step (A) is constituted by a plurality of sub-steps, it is also advisable to often extract the weakly adsorbing part in the plurality of sub-steps.

In the method of the present invention, it is preferable to use 3 or more types of eluents with different desorption powers, more preferably to use 4 or more types of eluents with different desorption powers, and more preferably to use the same desorption powers. 4 to 6 types of different eluents, especially 4 or 5 types of eluents with different desorption powers. The type of eluent is not particularly limited, and is appropriately set according to the relationship between the type of adsorbent and the type of components in the stock solution. For example, when an ion exchange resin is used as an adsorbent, the salt concentration of the eluent can be changed to prepare a plurality of eluents with different desorption capabilities. For example, when a cation exchange resin is used, a plurality of types of eluents with different NaCl concentrations can be used as two or more types of eluents.

The preferred embodiment of the combination of the sub-steps in the above step (A) is described below. The implementation of these aspects can be implemented using the system of FIG. 1 or a system that can be implemented in accordance with the system of FIG. 1. In addition, the following embodiment is an example of the present invention. For example, from the viewpoint of relative desorption force, two or more types of products that are given the status of the following eluent d-1 can also be prepared. Eluent, and during the substeps of supplying eluent d-1, change the type of eluent d-1 used. In this regard, the same applies to the eluents d-II to d-V. That is, in the present invention, when it is called "eluent d-1" in a certain embodiment, if in this embodiment, the "eluent d-1" is used in a different substep. , The "eluent d-1" used in the different substeps may be the same as each other or different from each other. In this regard, the same applies to the eluents d-II to d-V.

－Implementation style 1－ In Embodiment 1, a circulation system having 4 or more unit packed towers is used. Then, it is assumed that "the circulation system is divided into four sections 1 to 4 connected in a ring shape from the upstream side to the downstream side so that each section has at least one unit packed tower." In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In this embodiment 1, the following sub-steps (A1-1), (A2-1), and (A3-1) are executed in step (A). In the present invention, the so-called "execute sub-steps X, Y, and Z in step (A)" means that step (A) includes sub-steps X, Y, and Z, and the execution order of sub-steps X, Y, and Z , Can be appropriately set within the range that does not impair the efficacy of the present invention. In addition, step (A) may also include substeps X, Y, and Z other substeps. Regarding the aspect of "implementing sub-steps X, Y, and Z in step (A)", typically, the aspect in which sub-steps X, Y, and Z are executed in sequence as step (A) can be enumerated, but the present invention does not Limited to this aspect. That is, the so-called "perform substeps X, Y, and Z in step (A)" is not limited to "perform substep Y after substep X, perform substep Z after substep Y, and perform substep Z after substep Z. Step (B), perform sub-step X" after step (B). For example, in the above-mentioned aspect that includes other sub-steps, you can precede sub-step X [between step (B) and sub-step X], between sub-steps X and Y, between sub-steps Y and Z, and In at least one place between step Z and step (B), other sub-steps (sub-steps X, Y, and sub-steps other than sub-steps X, Y, and Z) are combined within the scope that does not impair the efficacy of the present invention. Sometimes by adjusting the supply flow rate, the flow rate, the strength of the eluent, etc., even if additional sub-steps other than the sub-steps X, Y, and Z are inserted, the desired effect can still be obtained. It's easy to understand this point.

＜Substep (A1-1)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Extract the strongly adsorbing part from the suction port C, use the upstream end of the section 2 as the eluent supply port D-II, and supply the eluent d-II from the eluent supply port D-II, and use the section 3 The upstream end of the section 4 is used as the raw liquid supply port F, the raw liquid is supplied from the raw liquid supply port F, the downstream end of the section 4 is used as the weakly adsorptive part extraction port A, and the weakly adsorbable part is extracted from the extraction port A, thereby making The desorption force of the eluent flowing in zone 1 is the strongest, and the desorption force of the eluent flowing in zone 2 is weaker than that of the eluent flowing in zone 1, making zones 3 and 4 circulate The desorption force of the eluent is weaker than the desorption force of the eluent circulating in zone 2. In the sub-step (A1-1), the above-mentioned supply of each liquid and the extraction of each part are continuously performed (that is, in the sub-step (A1-1), the above-mentioned supply of each liquid and the extraction of each part All are carried out often and without interruption). This point is the same in each sub-step described later. In addition, in the present invention and in this specification, the strength of the desorption force of the eluate described in each sub-step refers to the strength of the desorption force of the eluate in the sub-step, and it does not illustrate the difference in the sub-step. The strength of the desorption force of the eluent. For example, when it is explained that the desorption force of the eluent flowing in section 1 is the strongest in one sub-step constituting step (A), and it is also explained that in another sub-step constituting step (A), When the desorption force of the circulating eluent is the strongest, the desorption force of the eluent circulating in section 1 in this one sub-step is compared with the desorption of the eluent circulating in section 1 in the other sub-step The force can be the same or different.

＜Sub-step (A2-1)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The upstream end of the section 3 serves as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the suction port A, thereby making the section The desorption force of the eluent flowing in 1 is the strongest. The desorption force of the eluent flowing in section 2 is weaker than that of the eluent flowing in section 1. The desorption force of the eluent is weaker than the desorption force of the eluent flowing in section 2. The eluent supply port D-III in this substep (A2-1) is provided in the same piping as the stock solution supply port F in the above substep (A1-1).

<Sub-step (A3-1)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of section 3 serves as the middle adsorptive part extraction port B. The middle adsorptive part is extracted from the suction port B, and the upstream end of section 4 is used as the eluent supply port D-IV, from which eluent supply port D-IV supplies the eluent d-IV, and extracts the weakly adsorbing part from the extraction port A, thereby maximizing the desorption force of the eluent flowing in zone 1 and making the eluent flowing in zones 2 and 3 strongest The desorption force of the liquid is weaker than the desorption force of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 4 is stronger than that of the eluent flowing in zones 2 and 3. weak.

In the above substep (A2-1), the desorption force of the eluent flowing in the interval 3 and 4 should also be the same as that of the eluent flowing in the interval 4 in the above substep (A3-1). the same.

As an example of the above sub-step (A1-1), a sub-step of implementing the following sub-step (A1-1ex) can be cited, but the above-mentioned sub-step (A1-1) is not limited to the sub-step (A1-1ex). ＜Substep (A1－1ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strongly adsorbent part is drawn from the outlet C. The strongly adsorbed part is drawn from the outlet C. The upstream end of the section 2 is used as the eluent supply port D2, and the eluent supply port D2 supplies 4 types of eluents The eluent d2 with the second strongest desorption power of the zone 3 is used as the stock solution supply port F at the upstream end of the zone 3, the stock solution is supplied from the stock solution supply port F, and the downstream end of the zone 4 is taken as the weakly adsorbing part extraction port A , Extract the weakly adsorbing part from the extraction port A.

As an example of the above-mentioned sub-step (A2-1), a sub-step of implementing the following sub-step (A2-1ex) can be cited, but the above-mentioned sub-step (A2-1) is not limited to the sub-step (A2-1ex). ＜Substep (A2－1ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent d2 is supplied from the eluent supply port D2 to the upstream of zone 3 The side end is used as the eluent supply port D3. The eluent d3 with the weakest desorption force among the four eluents is supplied from the eluent supply port D3, and the weakly adsorbed portion A is drawn from the weakly adsorbing part. The sexual part. The eluent supply port D3 in this substep (A2-1ex) is provided in the same pipe as the stock solution supply port F in the above substep (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 cited, but the above-mentioned sub-step (A3-1) is not limited to the sub-step (A3-1ex). <Substep (A3-1ex)> The eluent d1 is supplied from the eluent supply port D1, the strongly absorptive portion is drawn from the strongly adsorbent portion extraction port C, and the eluent d2 is supplied from the eluent supply port D2 to the downstream side of the zone 3. The end is used as the suction port B for the middle adsorptive part. The middle adsorptive part is extracted from the suction port B. The upstream end of section 4 is used as the eluent supply port D4, and four types of eluent are supplied from the eluent supply port D4. Among the eluates d4 with the third strongest desorption force, the weakly adsorbable portion is extracted from the weakly adsorbable portion extraction port A.

Taking the state of having one unit packed tower in each section as an example, the above step (A) is shown in the flow chart of the state in which the above steps (A1-1ex), (A2-1ex) and (A3-1ex) are executed in sequence于图2。 In Figure 2. In Figure 2, the four-corner box represents a unit packing tower, and the number in the box represents the number of the unit packing tower (the numbers are attached in order from the left). After the step (A) of the above sub-steps (A1-1ex), (A2-1ex) and (A3-1ex) is completed in sequence, the original solution supply port F and the eluent supply port are set through step (B) D. The suction port A for the weakly adsorptive part, the suction port B for the medium adsorptive part and the suction port C for the strong adsorptive part move to the downstream side while maintaining their relative positional relationship, and then the above will be executed in order The flow chart of the sub-steps (A1-1ex), (A2-1ex) and (A3-1ex) is shown in Figure 3. The unit packed towers arranged in each section shown in Fig. 2 are displaced one by one to the downstream side in Fig. 3. At this time, "start with step (A) shown in Figure 2, and then execute step (B)" is regarded as a set of steps, and by implementing four sets of steps, it will return to the state shown in Figure 2 again.

-Implementation style 2- The second embodiment is the same as the first embodiment, using a circulation system with 4 or more unit packed towers. Then, it is assumed that "the circulation system is divided into four sections 1 to 4 connected in a ring shape from the upstream side to the downstream side so that each section has at least one unit packed tower." In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In this embodiment 2, the following sub-steps (A1-2), (A2-2), and (A3-2) are executed in order as step (A).

＜Substep (A1-2)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of section 3 is used as the stock solution supply port F. The raw liquid supply port F supplies raw liquid. The downstream end of section 4 is used as the weakly adsorbent part suction port A, and the weakly adsorbable part is drawn out from the suction port A, whereby the eluent flowing in sections 1 and 2 is removed. The adhesion force is the strongest, making the desorption force of the eluent flowing in sections 3 and 4 weaker than that of the eluent flowing in sections 1 and 2.

<Sub-step (A2-2)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Extract the strongly adsorbing part from the suction port C, use the upstream end of the section 2 as the eluent supply port D-II, and supply the eluent d-II from the eluent supply port D-II, and use the section 3 The upstream end of the eluate is used as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the zone 1 The desorption force of the circulating eluent is the strongest, making the desorption force of the eluent circulating in section 2 weaker than the desorbing force of the eluent circulating in section 1, so that the desorbing force in sections 3 and 4 The desorption force of the liquid is weaker than the desorption force of the eluent flowing in section 2. The eluent supply port D-I in this sub-step (A2-2) is provided in the same pipe as the eluent supply port D-II in the above-mentioned sub-step (A1-2). In addition, the eluent supply port D-III is provided in the same pipe as the stock solution supply port F. In this substep (A2-2), the desorption force of the eluent flowing in section 1 should be stronger than the desorption force of the eluent flowing in section 1 in the above substep (A1-2).

<Substep (A3-2)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-2) For this eluent d-II, the downstream end of the section 3 is used as the middle adsorptive part extraction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 4 is used as the eluent supply port D- IV, the eluent d-IV is supplied from the eluent supply port D-IV, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and The desorption force of the eluent flowing in zones 2 and 3 is weaker than that of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 4 is higher than that of zones 2 and 3. The desorption force of the solution is weaker.

As an example of the above-mentioned sub-step (A1-2), a sub-step of implementing the following sub-step (A1-2ex) can be cited, but the above-mentioned sub-step (A1-2) is not limited to the sub-step (A1-2ex). ＜Substep (A1－2ex)＞ The upstream end of section 1 is used as the eluent supply port D2, and the eluent d2 with the second strongest desorption force among the four eluents is supplied from the eluent supply port D2, and the upstream side of section 3 The end is used as a raw liquid supply port F, the raw liquid is supplied from the raw liquid supply port F, the downstream end of the section 4 is used as a weakly adsorbent portion suction port A, and the weakly adsorbable portion is drawn from the suction port A.

As an example of the aforementioned sub-step (A2-2), a sub-step of implementing the following sub-step (A2-2ex) can be cited, but the aforementioned sub-step (A2-2) is not limited to the sub-step (A2-2ex). ＜Substep (A2－2ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strongly adsorbent part is drawn from the outlet C. The strongly adsorbed part is drawn from the outlet C. The upstream end of the section 2 is used as the eluent supply port D2, and the eluent supply port D2 supplies 4 types of eluents The eluent d2, which has the second strongest desorption power, uses the upstream end of zone 3 as the eluent supply port D3. The eluent supply port D3 has the weakest desorption power among the four eluents. The eluted liquid d3 of, the weakly adsorbent part is drawn out from the weakly adsorbent part extraction port A. The eluent supply port D1 in this substep (A2-2ex) is provided in the same pipe as the eluent supply port D2 in the above substep (A1-2ex). In addition, the eluent supply port D3 is provided in the same piping as the stock solution supply port F in the above substep (A1-2ex).

As an example of the above-mentioned sub-step (A3-2), a sub-step of implementing the following sub-step (A3-2ex) can be cited, but the above-mentioned sub-step (A3-2) is not limited to the sub-step (A3-2ex). ＜Substep (A3－2ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent is supplied from the eluent supply port D2 in the substep (A2-2ex). For the eluent d2, the downstream end of section 3 is used as the middle adsorptive part extraction port B, the middle adsorptive part is extracted from the suction port B, and the upstream end of section 4 is used as the eluent supply port D4, from which the eluent The liquid supply port D4 supplies the eluent d4 with the third strongest desorption force among the four types of eluents, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the state of having one unit packed tower in each section as an example, the above step (A) is shown in the flow chart of the state in which the above substeps (A1-2ex), (A2-2ex) and (A3-2ex) are carried out in sequence于图4。 In Figure 4. In FIG. 4, the four-corner enclosure represents a unit packed tower of 1 unit, and the number in the enclosure represents the number of the unit packed tower. After the step (A) of the above sub-steps (A1-2ex), (A2-2ex) and (A3-2ex) is performed in sequence, the original solution supply port F and the eluent supply port D are set through step (B) , The extraction port A for the weakly adsorbing part, the pumping port B for the middle adsorbing part, and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －2ex), (A2－2ex) and (A3－2ex) flow charts are shown in Figure 5. The unit packed towers arranged in each section shown in Fig. 4 are displaced one by one to the downstream side in Fig. 5. At this time, "start with step (A) shown in FIG. 4, and then execute step (B)" is regarded as a set of steps, and by executing the 4 sets, it will return to the state shown in FIG. 4 again.

-Implementation style 3- Implementation aspect 3 uses a circulation system with more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into five sections 1 to 5 connected in an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In this embodiment 3, the following sub-steps (A1-3), (A2-3), and (A3-3) are sequentially performed as step (A).

<Sub-step (A1-3)> The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of section 3 is used as the stock solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 is used as the eluent supply port D-III, the eluent solution d-III is supplied from the eluent supply port D-III, and the downstream end of section 5 is taken as The weakly adsorptive part is drawn from the outlet A, and the weakly adsorptive part is extracted from the outlet A, thereby maximizing the desorption force of the eluent flowing in zones 1 and 2, and desorbing the eluent flowing in zone 3 The adhesion force is the same as or weaker than the desorption force of the eluent flowing in zones 1 and 2, so that the desorption of the eluent flowing in zones 4 and 5 The adhesion force is weaker than the desorption force of the eluent flowing in section 3.

＜Substep (A2－3)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Extract the strongly adsorbing portion from the suction port C, use the upstream end of the zone 2 as the eluent supply port D-II, and supply the eluent d-II from the eluent supply port D-II, and from the eluent The eluent supply port D-III supplies the eluent liquid d-III, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent liquid circulating in section 1 is maximized, and sections 2 and 3 The desorption force of the circulating eluent is weaker than the desorption force of the eluent circulating in section 1, so that the desorption force of the eluent circulating in sections 4 and 5 is higher than that of the eluent circulating in sections 2 and 3. The desorption force of the liquid is weaker. In this substep (A2-3), the desorption force of the eluent flowing in section 1 should be stronger than that of the eluent flowing in section 1 in the above substep (A1-3). The eluent supply port D-I in this substep (A2-3) is provided in the same pipe as the eluent supply port D-II in the above substep (A1-3).

＜Substep (A3－3)＞ The eluent d-I is supplied from the eluent supply port D-1, the strongly adsorbing part is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-3) For this eluent d-II, the downstream end of section 4 is used as the middle adsorptive part extraction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of section 5 is used as the eluent supply port D- IV, the eluent d-IV is supplied from the eluent supply port D-IV, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and The desorption force of the eluent flowing in zones 2, 3 and 4 is weaker than that of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 5 is higher than that of zones 2, 3 And the desorption force of the eluent flowing in 4 is weaker.

As an example of the above substep (A1-3), the following substep (A1-3ex) can be enumerated, but the above substep (A1-3) is not limited to the substep (A1-3ex). ＜Sub-step (A1-3ex)＞ The upstream end of section 1 is used as the eluent supply port D2, and the eluent d2 with the second strongest desorption force among the four eluents is supplied from the eluent supply port D2, and the upstream side of section 3 The end is used as the stock solution supply port F. The stock solution is supplied from the stock solution supply port F. The upstream end of the section 4 is used as the eluent supply port D3. The eluent supply port D3 supplies desorption among the four eluents. The eluent d3 with the weakest force is drawn with the weakly adsorbent portion at the downstream end of the section 5 as the weakly adsorbent portion A, and the weakly adsorbable portion is drawn from the pumped hole A.

As an example of the above-mentioned sub-step (A2-3), a sub-step of implementing the following sub-step (A2-3ex) can be cited, but the above-mentioned sub-step (A2-3) is not limited to the sub-step (A2-3ex). ＜Substep (A2－3ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strongly adsorbent part is drawn from the outlet C. The strongly adsorbed part is drawn from the outlet C. The upstream end of the section 2 is used as the eluent supply port D2, and the eluent supply port D2 supplies 4 types of eluents The eluent d2 with the second strongest desorption power of, is supplied from the eluent supply port D3, and the weakly adsorbable portion is extracted from the weakly adsorbent portion extraction port A. The eluent supply port D1 in this substep (A2-3ex) is provided in the same pipe as the eluent supply port D2 in the above substep (A1-3ex).

As an example of the above substep (A3-3), the following substep (A3-3ex) can be enumerated, but the above substep (A3-3) is not limited to the substep (A3-3ex). ＜Substep (A3－3ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent is supplied from the eluent supply port D2 in the substep (A2-3ex). For the eluent d2, the downstream end of the section 4 is used as the middle adsorptive part suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D4. The liquid supply port D4 supplies the eluent d4 with the third strongest desorption force among the four types of eluents, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the state of having one unit packed tower in each section as an example, the flow chart showing the state of performing the above steps (A) in order of the above sub-steps (A1-3ex), (A2-3ex) and (A3-3ex) In Figure 6. In FIG. 6, the four-corner enclosing frame represents a unit packing tower of 1 unit, and the number in the enclosure represents the number of the unit packing tower. After the step (A) of the above sub-steps (A1-3ex), (A2-3ex) and (A3-3ex) is performed in sequence, the original solution supply port F and the eluent supply port D are set through step (B) , The pumping outlet A for the weakly adsorbing part, the pumping outlet B for the middle adsorbing part, and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －3ex), (A2－3ex) and (A3－3ex) flow charts are shown in Figure 7. The unit packed towers arranged in each section shown in Fig. 6 are displaced one by one to the downstream side in Fig. 7. At this time, "start with step (A) shown in FIG. 6, and then execute step (B)" is regarded as a set of steps, and by executing 5 sets of steps, it will return to the state shown in FIG. 6 again.

-Implementation style 4- Implementation aspect 4 uses a circulation system with more than 7 unit packed towers. Then, it is assumed that "the circulation system is divided into five sections 1 to 5 connected in an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. In addition, five types of eluents d-I to d-V with different desorption powers were used as eluents. In this embodiment 4, the following sub-steps (A1-4), (A2-4), and (A3-4) are sequentially performed as step (A).

＜Substep (A1-4)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of section 3 is used as the stock solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 is used as the eluent supply port D-III, the eluent solution d-III is supplied from the eluent supply port D-III, and the downstream end of section 5 is taken as The extraction port A extracts the weakly adsorbing part from the extraction port A, whereby the desorption force of the eluent flowing in sections 1 and 2 is the strongest, and the desorption force of the eluent flowing in section 3 is equal to The desorption power of the eluent flowing in zones 1 and 2 is the same or weaker than that of the eluent flowing in zones 1 and 2. The desorption force of the eluent flowing in zones 4 and 5 is higher than The desorption force of the eluent flowing in section 3 is weaker.

＜Substep (A2－4)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Extract the strongly adsorbing part from the suction port C, use the upstream end of the section 2 as the eluent supply port D-II, and supply the eluent d-II from the eluent supply port D-II, and use the section 4 The upstream end of the eluate is used as the eluent supply port D-IV, the eluent d-IV is supplied from the eluent supply port D-IV, and the weakly adsorptive portion is extracted from the extraction port A, thereby making the zone 1 The desorption force of the circulating eluate is the strongest. The desorption force of the eluate circulating in sections 2 and 3 is weaker than that of the eluent circulating in section 1. The desorption force of the eluent is weaker than the desorption force of the eluent flowing in sections 2 and 3. In this substep (A2-4), the desorption force of the eluent flowing in section 1 should be stronger than the desorption force of the eluent flowing in section 1 in the above substep (A1-4). The eluent supply port D-I in this substep (A2-4) is provided in the same pipe as the eluent supply port D-II in the above substep (A1-4).

<Substep (A3-4)> The eluent d-1 is supplied from the eluent supply port D-1, the strongly adsorbent portion is extracted from the suction port C, and the eluent is supplied from the eluent supply port D2 in the substep (A2-4). For the eluent d2, the downstream end of the zone 4 is used as the middle adsorptive part extraction port B, the middle adsorptive part is drawn from the extraction port B, and the upstream end of the zone 5 is used as the eluent supply port DV from The eluent supply port D-V supplies the eluent d-V, and the weakly adsorptive part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in the section 1 is maximized, and the sections 2, 3 And the desorption force of the eluent flowing in 4 is weaker than the desorption force of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zone 5 is higher than that of zones 2, 3 and 4. The desorption force of the solution is weaker.

As an example of the above sub-step (A1-4), a sub-step of implementing the following sub-step (A1-4ex) can be cited, but the above-mentioned sub-step (A1-4) is not limited to the sub-step (A1-4ex). ＜Substep (A1－4ex)＞ The upstream end of section 1 is used as the eluent supply port D2, and the eluent d2 with the second strongest desorption force among the five eluents is supplied from the eluent supply port D2, and the upstream side of section 3 The end 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 section 4 is used as the eluent supply port D3, and the eluent supply port D3 supplies desorption among the five eluents. The eluent d3 with the weakest force is drawn with the weakly adsorbent portion at the downstream end of the section 5 as the weakly adsorbent portion A, and the weakly adsorbed portion is drawn from the pumped hole A.

In the above substep (A3-3), the desorption capacity of the eluent flowing in the sections 4 and 5 should also be the same as the desorption force of the eluent flowing in the section 5 in the above substep (A3-4). the same.

As an example of the above-mentioned sub-step (A2-4), a sub-step of implementing the following sub-step (A2-4ex) can be cited, but the above-mentioned sub-step (A2-4) is not limited to the sub-step (A2-4ex). ＜Substep (A2－4ex)＞ The upstream end of section 1 is used as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the five eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The highly absorbent part is drawn out from the outlet C, the highly absorbent part is drawn out from the outlet C, and the upstream end of the section 2 is used as the eluent supply port D2, and the eluent supply port D2 supplies 5 kinds of eluents The eluent d2, which has the second strongest desorption power, uses the upstream end of zone 4 as the eluent supply port D4, and the eluent supply port D4 supplies the fourth desorption power among the five eluent The strong eluent d4 draws out the weakly adsorbent part from the weakly adsorbent part extraction port A. The eluent supply port D1 in this substep (A2-4ex) is provided in the same pipe as the eluent supply port D2 in the above substep (A1-4ex).

As an example of the above sub-step (A3-4), a sub-step of implementing the following sub-step (A3-4ex) can be cited, but the above-mentioned sub-step (A3-4) is not limited to the sub-step (A3-4ex). ＜Substep (A3－4ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent is supplied from the eluent supply port D2 in the substep (A2-4ex). For the eluent d2, the downstream end of the section 4 is used as the middle adsorptive part suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D5. The liquid supply port D5 supplies the eluent d5 with the third strongest desorption force among the five eluent liquids, and the weakly adsorptive portion is extracted from the weakly adsorptive portion extraction port A.

An example of a flow in which the above-mentioned step (A) sequentially executes the above-mentioned sub-steps (A1-4ex), (A2-4ex), and (A3-4ex) is shown in FIG. 8. In FIG. 8, the four-corner enclosing frame represents a unit packing tower of 1 unit, and the number in the enclosing frame represents the number of the unit packing tower. In addition, the aspect shown in Fig. 8 has 7 unit packed towers, one unit packed tower is included in zone 1, two unit packed towers are included in zone 2, two unit packed towers are included in zone 3, and zone 4 is included 1 unit packed tower includes 1 unit packed tower in section 5. After the step (A) of the above sub-steps (A1-4ex), (A2-4ex) and (A3-4ex) is completed in sequence, through step (B), make the stock solution supply port F and the eluent supply port D , The extraction port A for the weakly adsorbing part, the pumping port B for the middle adsorbing part and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －4ex), (A2－4ex) and (A3－4ex) flow charts are shown in Figure 9. The unit packed towers arranged in each section shown in Fig. 8 are each displaced by 1 unit to the downstream side in Fig. 9. At this time, "start with step (A) shown in FIG. 8, and then execute step (B)" is regarded as a set of steps, and by executing 7 sets of them, it returns to the state shown in FIG. 8 again.

-Implementation style 5- Implementation aspect 5 uses a circulation system with more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into five sections 1 to 5 connected in an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In this embodiment 5, the following sub-steps (A1-5), (A2-5), and (A3-5) are sequentially performed as step (A).

<Substep (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 eluent supply port D-III, and the eluent supply port D-III is supplied For the eluent d-III, the downstream end of the section 5 is used as the weakly adsorbing part extraction port A, and the weakly adsorbing part is extracted from the pumping port A, thereby increasing the desorption power of the eluent flowing in the section 3 The strongest is to make the desorption force of the eluent flowing in sections 4 and 5 weaker than the desorption force of the eluent flowing in section 3.

＜Substep (A2－5)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Extract the strongly adsorbing portion from the suction port C, use the upstream end of the zone 2 as the eluent supply port D-II, and supply the eluent d-II from the eluent supply port D-II, and from the eluent The eluent supply port D-III supplies the eluent liquid d-III, and the weakly adsorbable part is extracted from the extraction port A, whereby the desorption force of the eluent liquid circulating in section 1 is maximized, and sections 2 and 3 The desorption force of the circulating eluent is weaker than the desorption force of the eluent circulating in section 1, so that the desorption force of the eluent circulating in sections 4 and 5 is higher than that of the eluent circulating in sections 2 and 3. The desorption force of the liquid is weaker.

＜Substep (A3－5)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of section 4 serves as the middle adsorptive part extraction port B. The middle adsorptive part is extracted from the suction port B, and the upstream end of section 5 is used as the eluent supply port D-IV from the eluent supply port D-IV supplies the eluent d-IV, and extracts the weakly adsorptive part from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and the desorbing power in sections 2, 3, and 4 is maximized. The desorption force of the eluent is weaker than the desorption force of the eluent flowing in zone 1, and the desorption force of the eluent flowing in zone 5 is higher than that of the eluent flowing in zones 2, 3, and 4. The desorption force is weaker. In this substep (A3-5), the desorption force of the eluent flowing in section 1 should be the same as that of the eluent flowing in section 1 in the above substep (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 cited, but the above-mentioned sub-step (A1-5) is not limited to the sub-step (A1-5ex). ＜Substep (A1－5ex)＞ 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. The upstream end of the section 4 is used as the eluent supply port D3, and four types of eluents are supplied from the eluent supply port D3 The eluent d3 with the weakest desorption force among the liquids is drawn out of the weakly adsorbing portion A at the downstream end of the section 5, and the weakly adsorbing portion is drawn out from the pumping port A.

As an example of the above substep (A2-5), the following substep (A2-5ex) can be enumerated, but the above substep (A2-5) is not limited to the substep (A2-5ex). ＜Substep (A2－5ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strongly adsorbent part is drawn from the outlet C. The strongly adsorbed part is drawn from the outlet C. The upstream end of the section 2 is used as the eluent supply port D2, and the eluent supply port D2 supplies 4 types of eluents The eluent d2 with the second strongest desorption power of, is supplied from the eluent supply port D3, and the weakly adsorbable portion is extracted from the weakly adsorbent portion extraction port A.

As an example of the above-mentioned sub-step (A3-5), the following sub-step (A3-5ex) can be cited, but the above-mentioned sub-step (A3-5) is not limited to the sub-step (A3-5ex). ＜Substep (A3－5ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent d2 is supplied from the eluent supply port D2 to the downstream of zone 4 The end of the side is used as the middle adsorptive part extraction port B. The middle adsorptive part is extracted from the outlet B. The upstream end of section 5 is used as the eluent supply port D4, and four types of eluents are supplied from the eluent supply port D4 The eluent d4 with the third strongest desorption force in the liquid is drawn out from the weakly adsorbent part extraction port A.

Taking the state of having one unit packed tower in each section as an example, the above step (A) is shown in the flow chart of the state in which the above sub-steps (A1-5ex), (A2-5ex) and (A3-5ex) are executed in sequence于图13。 In Figure 13. In FIG. 13, the four-corner enclosing frame indicates a unit packing tower of 1 unit, and the number in the enclosure indicates the number of the unit packing tower. After step (A) of the above sub-steps (A1-5ex), (A2-5ex) and (A3-5ex) are performed in sequence, step (B) is used to make the stock solution supply port F and the eluent supply port D , The extraction port A for the weakly adsorbing part, the pumping port B for the middle adsorbing part, and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －5ex), (A2－5ex) and (A3－5ex) flow charts are shown in Figure 14. The unit packed towers arranged in each section shown in Fig. 13 are displaced one by one to the downstream side in Fig. 14. At this time, "start with step (A) shown in FIG. 13, and then execute step (B)" is regarded as a set of steps, and by executing five sets of steps, it will return to the state shown in FIG. 13 again.

-Implementation style 6- Implementation aspect 6 uses a circulation system with more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into five sections 1 to 5 connected in an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In the sixth embodiment, the following sub-steps (A1-6), (A2-6), and (A3-6) are executed in order as step (A).

＜Substep (A1－6)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the downstream end of section 3 is used as the middle adsorptive part extraction port B , Withdraw the middle adsorptive part from the suction port B, use the upstream end of the section 4 as the eluent supply port D-IV, and supply the eluent d-IV from the eluent supply port D-IV, and use the section 5 The downstream end of the A is used as the weakly adsorbing part extraction port A, and the weakly adsorbing part is extracted from the pumping port A, whereby the desorption force of the eluent flowing in the sections 1, 2 and 3 is maximized, and the sections 4 and 5 The desorption force of the eluent flowing through is weaker than the desorption force of the eluent flowing through zones 1, 2 and 3.

<Substep (A2-6)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Withdraw the strongly adsorbing portion from the suction port C, use the upstream end of the section 3 as the stock solution supply port F, supply the stock solution from the stock solution supply port F, and use the upstream end of the section 4 as the eluent supply port D- III. The eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized. The desorption force of the eluent flowing in zone 3 is weaker than that of the eluent flowing in zone 1, so that the desorption force of the eluent flowing in zones 4 and 5 is higher than that of the eluent flowing in zone 3. The desorption force of the liquid is weaker. In this substep (A2-6), the desorption force of the eluent flowing in section 1 should be stronger than that of the eluent flowing in section 1 in the above substep (A1-6). The eluent supply port D-1 in this sub-step (A2-6) is provided in the same pipe as the eluent supply port D-II in the aforementioned substep (A1-6).

<Substep (A3-6)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the interval 1 The desorption force of the eluent flowing in 1 is the strongest, and the desorption force of the eluent flowing in sections 2 and 3 is weaker than that of the eluent flowing in section 1. The desorption force of the circulating eluate is weaker than the desorption force of the circulating eluate in sections 2 and 3. In this sub-step (A3-6), the desorption force of the eluent flowing in section 1 should be the same as that of the eluent flowing in section 1 in the above sub-step (A2-6).

As an example of the above-mentioned sub-step (A1-6), the following sub-step (A1-6ex) can be enumerated, but the above-mentioned sub-step (A1-6) is not limited to the sub-step (A1-6ex). ＜Substep (A1－6ex)＞ The upstream end of section 1 is used as the eluent supply port D2, and the eluent d2 with the second strongest desorption force among the four eluents is supplied from the eluent supply port D2, and the downstream side of section 3 The end is used as the suction port B for the middle adsorptive part. The middle adsorptive part is extracted from the suction port B. The upstream end of section 4 is used as the eluent supply port D4, and four types of eluent are supplied from the eluent supply port D4. Among the eluates d4 with the third strongest desorption force, the downstream end of the section 5 is taken as the weakly adsorbing part extraction port A, and the weakly adsorbing part is extracted from the pumping port A.

As an example of the above-mentioned sub-step (A2-6), a sub-step of implementing the following sub-step (A2-6ex) can be cited, but the above-mentioned sub-step (A2-6) is not limited to the sub-step (A2-6ex). ＜Substep (A2－6ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strong adsorbent part is drawn from the outlet C. The strong adsorbent part is drawn from the outlet C. The upstream end of the section 3 is used as the stock solution supply port F, the stock solution is supplied from the stock supply port F, and the upstream end of the zone 4 is used as the melt The eluent supply port D3 supplies the eluent d3 with the weakest desorption force among the four types of eluents from the eluent supply port D3, and extracts the weakly adsorbent portion from the weakly adsorbent portion 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 cited, but the above-mentioned sub-step (A3-6) is not limited to the sub-step (A3-6ex). ＜Sub-step (A3－6ex)＞ The eluent d1 is supplied from the eluent supply port D1, and the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C. The upstream end of the zone 2 is used as the eluent supply port D2, and the eluent The supply port D2 supplies the eluent solution d2, the eluent solution d3 is supplied from the eluent solution supply port D3, and the weakly adsorbent portion is extracted from the weakly adsorbent portion extraction port A.

Taking the state of having one unit packed tower in each section as an example, the above step (A) is shown in the flow chart of the state in which the above sub-steps (A1-6ex), (A2-6ex) and (A3-6ex) are executed in sequence于图15。 In Figure 15. In FIG. 15, the four-corner frame represents a unit packed tower of 1 unit, and the number in the frame indicates the number of the unit packed tower. After step (A) of the above substeps (A1-6ex), (A2-6ex) and (A3-6ex) are performed in sequence, step (B) is used to make the stock solution supply port F and the eluent supply port D , The extraction port A for the weakly adsorbing part, the pumping port B for the middle adsorbing part, and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －6ex), (A2－6ex) and (A3－6ex) flow charts are shown in Figure 16. The unit packed towers arranged in each section shown in Fig. 15 are displaced one by one to the downstream side in Fig. 16. At this time, "start with step (A) shown in FIG. 15, and then execute step (B)" is regarded as a set of steps, and by executing 5 sets of steps, it will return to the state shown in FIG. 15 again.

-Implementation style 7- Implementation aspect 7 uses a circulation system with more than 5 unit packed towers. Then, it is assumed that "the circulation system is divided into five sections 1 to 5 connected in an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. In addition, four types of eluents d-I to d-IV with different desorption power were used as eluents. In this embodiment 7, the following sub-steps (A1-7), (A2-7), and (A3-7) are sequentially performed as step (A).

＜Substep (A1-7)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-1, and the downstream end of section 1 is used as the suction port C for the strongly adsorbing portion , Withdraw the strongly adsorbing portion from the suction port C, use the upstream end of the section 3 as the stock solution supply port F, supply the stock solution from the stock supply port F, and use the upstream end of the section 4 as the eluent supply port D-III , The eluent d-III is supplied from the eluent supply port D-III, the downstream end of the section 5 is used as the suction port A, and the weakly adsorbable part is extracted from the suction port A, thereby making the section 1 The desorption force of the circulating eluent is the strongest, making the desorption force of the eluent circulating in section 3 weaker than that of the eluent circulating in section 1, so that the desorbing force in sections 4 and 5 The desorption force of the liquid is weaker than the desorption force of the eluent flowing in zone 3.

<Substep (A2-7)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby making the interval 1 The desorption force of the eluent flowing in 1 is the strongest, and the desorption force of the eluent flowing in sections 2 and 3 is weaker than that of the eluent flowing in section 1. The desorption force of the circulating eluate is weaker than the desorption force of the circulating eluate in sections 2 and 3.

<Substep (A3-7)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of section 4 serves as the middle adsorptive part extraction port B. The middle adsorptive part is extracted from the suction port B, and the upstream end of section 5 is used as the eluent supply port D-IV from the eluent supply port D-IV supplies the eluent d-IV, and extracts the weakly adsorptive part from the extraction port A, whereby the desorption force of the eluent flowing in section 1 is maximized, and the desorbing power in sections 2, 3, and 4 is maximized. The desorption force of the eluent is weaker than the desorption force of the eluent flowing in zone 1, and the desorption force of the eluent flowing in zone 5 is higher than that of the eluent flowing in zones 2, 3, and 4. The desorption force is weaker.

As an example of the above substep (A1-7), the following substep (A1-7ex) can be enumerated, but the above substep (A1-7) is not limited to the substep (A1-7ex). ＜Substep (A1－7ex)＞ The upstream end of section 1 is taken as the eluent supply port D1, and the eluent d1 with the strongest desorption force among the four types of eluents is supplied from the eluent supply port D1, and the downstream end of section 1 is taken as The strong adsorbent part is drawn from the outlet C. The strong adsorbent part is drawn from the outlet C. The upstream end of the section 3 is used as the stock solution supply port F, the stock solution is supplied from the stock supply port F, and the upstream end of the zone 4 is used as the melt The eluent supply port D3 is used to supply the eluent d3 with the weakest desorption force among the four eluents from the eluent supply port D3, and the downstream end of the section 5 is used as the weakly adsorbing part suction port A, From the extraction port A, the weakly adsorbable part is extracted.

As an example of the above substep (A2-7), the following substep (A2-7ex) can be enumerated, but the above substep (A2-7) is not limited to the substep (A2-7ex). ＜Substep (A2－7ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, the eluent d3 is supplied from the eluent supply port D3, and the weakly adsorbable portion Partial extraction port A extracts the weakly adsorbing part.

As an example of the above substep (A3-7), the following substep (A3-7ex) can be enumerated, but the above substep (A3-7) is not limited to the substep (A3-7ex). ＜Sub-step (A3－7ex)＞ The eluent d1 is supplied from the eluent supply port D1, the strongly adsorptive portion is extracted from the strongly adsorbent portion extraction port C, and the eluent d2 is supplied from the eluent supply port D2 to the downstream of zone 4 The end of the side is used as the middle adsorptive part extraction port B. The middle adsorptive part is extracted from the outlet B. The upstream end of section 5 is used as the eluent supply port D4, and four types of eluents are supplied from the eluent supply port D4 The eluent d4 with the third strongest desorption force in the liquid is drawn out from the weakly adsorbent part extraction port A.

Taking the state of having one unit packed tower in each section as an example, the above step (A) is shown in the flow chart of the state where the above steps (A1-7ex), (A2-7ex) and (A3-7ex) are executed in sequence于图17。 In Figure 17. In FIG. 17, the four-corner enclosing frame indicates a unit packing tower of 1 unit, and the number in the enclosure indicates the number of the unit packing tower. After step (A) of the above substeps (A1-7ex), (A2-7ex) and (A3-7ex) are performed in sequence, step (B) is used to set the stock solution supply port F and the eluent supply port D , The extraction port A for the weakly adsorbing part, the pumping port B for the middle adsorbing part, and the pumping outlet C for the strong adsorbing part move to the downstream side while maintaining their relative positional relationship. Then, the above sub-steps (A1 －7ex), (A2－7ex) and (A3－7ex) flow charts are shown in Figure 18. The unit packed towers arranged in each section shown in FIG. 17 are displaced one by one to the downstream side in FIG. 18. At this time, "start with step (A) shown in FIG. 17, and then execute step (B)" is regarded as a set of steps, and by executing 5 sets of steps, it will return to the state shown in FIG. 17 again.

In the method of the present invention, the supply of the target liquid to the target place, or the extraction of the target liquid from the target place, can be carried out by appropriately adjusting the operation of the pump installed at each part of the circulation system and the opening and closing of the valve at each part. It. That is, the method of supplying the target fluid or extracting the target part in the circulation system itself is conventionally known. In addition, the supply flow rate or the extraction flow rate of each liquid can also be appropriately set in accordance with the purpose of processing efficiency and the like.

In the method of the present invention, the purification target component can be any one of a strong adsorption component, a medium adsorption component, and a weak adsorption component, but it is a more suitable method for purifying the intermediate adsorption component. The method of the present invention can be applied to the purification of protein. Since the method of the present invention can obtain high-purity intermediate adsorptive components, it is a suitable method for obtaining high-purity target protein from a stock solution that contains decomposed products or aggregates in addition to the target protein. The above-mentioned protein is not particularly limited. For example, antibodies may be the components to be purified. In the present invention, the "antibody" may be a naturally-occurring antibody, a chimeric antibody, or an antibody fragmented by enzymes and the like [for example, F(ab') ₂ fragment, Fab' fragment, Fab fragment]. In addition, it also includes single-chain antibodies or diabody, triabody, or minibody. In addition, it may also be a single domain antibody. In addition, these substances are only examples, and proteins and their derivatives having specific binding power to antigens are all included in the concept of the antibody of the present invention. In the method of the present invention, the highly purified antibody can also be used as an antibody drug. That is, by applying the method of the present invention to extract the antibody contained in the stock solution, a method for manufacturing an antibody drug can be provided. More specifically, using the method of the present invention, the antibody drug can be obtained by extracting the antibody contained in the culture medium of the antibody-producing cells and/or the extract of the antibody-producing cells as the original liquid. In the present invention, the term "antibody-producing cell culture medium" or "antibody-producing cell extract" refers to a culture medium of antibody-producing cells or an extract of antibody-producing cells subjected to centrifugal separation or chromatography, etc. All kinds of processing, and the meaning of being in a certain degree of separation or even refinement.

In the method of the present invention, the adsorbent filled in the unit packed tower is appropriately selected according to the components to be purified, and various adsorbents can be used. For example, strong acid cation exchange resin, weak acid cation exchange resin, strong basic anion exchange resin, weakly basic anion exchange resin, synthetic adsorbent, zeolite, silica gel, and functional group bonded silica gel (preferably octadecyl silica Bonded silica gel). In addition, other gel filtration chromatography materials and affinity adsorption materials can be used as adsorbents. When the target component is protein, the adsorbent is preferably an ion exchange resin. Among them, cation exchange resin is more suitable.

The chromatographic separation system of the simulated moving layer mode of the present invention is a system for implementing the method of the present invention. That is, the simulated moving layer chromatography separation system of the present invention has the structure of the above-mentioned circulatory system, and the circulatory system is a system that can sequentially repeat the operation of step (A) and step (B). [Example]

Hereinafter, the present invention will be described in further detail based on examples, but the present invention is not limited to the following examples.

[Preparation of stock solution] After culturing cells that produce human immunoglobulin G2 (IgG2), the supernatant of the culture medium is desalted by dialysis, and the salt concentration adjusted by NaCl is added as the original solution. The content of antibodies, fragments, and aggregates contained in the supernatant is as follows. In the following table, fragment 1 is a protein contained in a portion with a molecular weight near 5000 and a molecular weight less than 25,000, and fragment 2 is a protein contained in a portion with a molecular weight of 25,000 or more and less than 50,000. In addition, the antibody is a protein contained in a portion having a molecular weight near 150,000 and having a molecular weight of 50,000 or more and less than 300,000. In addition, the aggregate is a protein contained in a portion having a molecular weight of 300,000 or more. The composition of the following components is determined by High Performance Liquid Chromatography (HPLC) using an analytical column (Tosoh TSKgel G3000SWXL).

[Table 1] Table 1 Ingredients in the clear solution above the culture solution Concentration (g/L) Fragment 1 0.02 Fragment 2 0.326 antibody 1.564 Aggregate 0.090

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

[Solution] Phosphate buffer solutions of various NaCl concentrations were prepared from the following A solution and B solution and used as an eluent. ＜A liquid＞ 20mM phosphate buffer, pH 6.0. ＜B liquid＞ 20mM phosphate buffer containing 0.3M (17.53g/L) NaCl concentration, pH 6.0.

[Comparative example 1] Single string, step gradient ＜Pipe string＞ 10mm diameter x 100mm length, 1 piece. ＜Original solution＞ The NaCl concentration in the stock solution is 2.05g/L. ＜Elute solution＞ Use the following eluent.

[Table 2] Table 2 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 2.39 D2 2.66 D3 17.53

<Operating conditions> Follow the steps 1 to 6 below in order. The flowchart of steps 1 to 6 is shown in FIG. 10. Under the following operating conditions, the recovery rate of fragment 1 and fragment 2 of the weakly adsorbing part is above 98%, the recovery rate of antibody in the middle-adsorbing part is above 98%, and the recovery rate of agglomerates of the strongly adsorbing part is above Above 98% condition. About this point, it is the same also in each comparative example and an Example mentioned later.

[Table 3] Table 3 step 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 Fragment 1 and Fragment 2 of the weakly adsorbing portion, the recovery rate of the antibody in the middle adsorbing portion, and the recovery rate of the aggregates of the strongly adsorbing portion are shown in the following table. The recovery rate was calculated by 100×[mass in part]/[mass in stock solution].

[Table 4] Table 4 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.2 Middle adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

－Separation treatment efficiency－ Based on each unit of adsorbent volume (unit: "L (liter)-R", R is the abbreviation of Resin), per unit time (unit: "h (hour)") stock solution processing volume (unit: "L ( Liter)-stock solution ") is the separation processing efficiency. In addition, in a multi-column system using a plurality of branch columns described below, the volume of the adsorbent is the total amount of adsorbent contained in all the columns. The separation treatment efficiency in Comparative Example 1 was 6.04 (L-stock solution)/(L-R)·h.

[Comparative example 2] Single string, step gradient ＜Pipe string＞ 10mm diameter×400mm length, 1 piece. ＜Original solution＞ The NaCl concentration in the stock solution is 2.05g/L. ＜Elute solution＞ Use the following eluent.

[Table 5] Table 5 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 2.35 D2 2.62 D3 17.53

<Operating conditions> Follow the steps 1 to 6 below in order. The flowchart of steps 1 to 6 is shown in Figure 10.

[Table 6] Table 6 step 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 Fragment 1 and Fragment 2 of the weakly adsorbing portion, the recovery rate of the antibody in the middle adsorbing portion, and the recovery rate of the aggregates of the strongly adsorbing portion are shown in the following table.

[Table 7] Table 7 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Comparative example 3] Multi-column, gradient, and simulated moving layer method ＜Pipe string＞ Diameter 10mm × length 100mm, 4 pieces. ＜Original solution＞ The concentration of NaCl in the stock solution is 2.23g/L. ＜Elute solution＞ Use the following eluent.

[Table 8] Table 8 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.28 D3 2.23

<Operating conditions> FIG. 11 shows a flowchart of the operation of Comparative Example 3. Taking the first to fourth steps shown in Fig. 11 as one cycle, 10 cycles are performed. Between each step, execute the original solution supply port F, the eluent supply port D (D1～D3), the weakly adsorbent part extraction port A, the medium adsorbent part pump out port B, and the strong adsorbent part pump out port C. While maintaining the relative positional relationship, move one pipe string to the downstream side. In addition, the first to fourth steps shown in FIG. 11 correspond to step (A) of the present invention, but different from this step (A), it is not composed of a plurality of sub-steps (in other words, one sub-step constitutes one step) ). "D1", "D2", and "D3" in FIG. 11 are all eluent supply ports, and eluent solutions D1, D2, and D3 are respectively supplied. "C" in Figure 11 is the extraction port for the strong adsorption part, and the strong adsorption part is extracted. Similarly, "B" is the suction port for the middle adsorptive part, and the middle adsorptive part is extracted, and "A" is the suction port for the weak adsorptive part, and the weak adsorptive part is extracted. The supply flow rates of the eluent (D1 to D3) and the stock solution (F) in each step shown in FIG. 11 are as follows. In addition, the following table does not record the flow rate of the liquid to be drawn out, but the flow rate of the strongly adsorptive portion drawn from the outlet C of the strongly adsorptive portion is the same as the flow rate of the eluent solution D1. In addition, the flow rate of the middle adsorptive portion extracted from the middle adsorptive portion suction port B is the same as the flow rate of the supply eluent D2. In addition, the flow rate of the weakly adsorbent portion extracted from the weakly adsorbent portion extraction port A is the sum of the flow rate of the eluent solution D3 supplied and the flow rate of the original solution supplied from the original solution supply port F. That is, the supply flow rate and the extraction flow rate are always the same, and this point is also the same in the following comparative examples and even examples.

[Table 9] Table 9 1 step time (minutes) 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 Fragment 1 and Fragment 2 of the weakly adsorbing portion, the recovery rate of the antibody in the middle adsorbing portion, and the recovery rate of the aggregates of the strongly adsorbing portion are shown in the following table.

[Table 10] Table 10 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 Over 99 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Comparative example 4] Multi-column, gradient, and simulated moving layer method ＜Pipe string＞ Diameter 10mm × length 100mm, 4 pieces. ＜Original solution＞ The concentration of NaCl in the stock solution is 2.24g/L. ＜Elute solution＞ Use the following eluent.

[Table 11] Table 11 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.26 D3 2.24

<Operating conditions> FIG. 12 shows a flowchart of the operation of Comparative Example 4. Take the first to fourth steps shown in Fig. 12 as one cycle and implement 10 cycles. In addition, each step shown in FIG. 12 is composed of two sub-steps, the first sub-step and the second sub-step. The second sub-step does not perform liquid supply and extraction, but circulates the fluid in the circulation system. The supply flow rates of the eluent (D1 to D3) and the stock solution (F) in each step shown in FIG. 12 are as follows.

[Table 12] Table 12 Sub-step time (minutes) Flow rate (mL/min) Substep 1 32.7 D1 1.90 D2 17.4 D3 1.33 F 3.82 Step 2 0.48 Loop 0.94

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 13] Table 13 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 Over 99 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Example 1] Multi-column, gradient, and simulated moving layer method ＜Pipe string＞ Diameter 10mm × length 100mm, 4 pieces. ＜Original solution＞ The concentration of NaCl in the stock solution is 1.93g/L. ＜Elute solution＞ Use the following eluent.

[Table 14] Table 14 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 3.59 D3 1.93 D4 2.21

<Operating conditions> Use the combination of the sub-steps shown in Figure 2 to form step (A). Taking this step (A) and the subsequent step (B) as one set of steps, 4 sets are executed as 1 cycle, and 10 cycles are implemented. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

[Table 15] Table 15 Sub-step time (minutes) Flow rate (mL/min) Substep (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-step (A3-1) 3.75 D1 13.87 D2 5.62 D4 15.21

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 16] Table 16 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorption part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Example 2] Multi-tubular column, gradient, simulated moving layer method <Tube column> Diameter 10mm×Length 100mm, 4 pieces. <The stock solution> The NaCl concentration in the stock solution is 2.02 g/L. <Eluent> The following eluent was used. [Table 17] Table 17 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 3.57 D3 2.02 D4 2.21

<Operating conditions> Use the combination of the sub-steps shown in Figure 4 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, and execute the 4 sets as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 19] Table 19 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive 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-tubular column, gradient, simulated moving layer method <Tube column> Diameter 10mm×Length 805mm, 5 pieces. <The stock solution> The concentration of NaCl in the stock solution is 2.46 g/L. <Eluent> The following eluent was used. [Table 20] Table 20 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.46 D3 0.00 D4 1.98

<Operating conditions> Use the combination of the sub-steps shown in Figure 6 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, execute 5 sets of them as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 22] Table 22 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive 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-tube column, gradient, and simulated moving layer method <Tube column> Diameter 10mm×Length 100mm, 7 pieces. <The stock solution> The concentration of NaCl in the stock solution is 2.57g/L. <Eluent> The following eluent was used. [Table 23] Table 23 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.57 D3 0.00 D4 0.37 D5 1.80

<Operating conditions> Use the combination of the sub-steps shown in Figure 8 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, execute 7 sets of them as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D5) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 25] Table 25 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

As mentioned above, it can be seen that in the chromatographic separation of the simulated moving layer method, two or more eluents are used, and the weakly adsorptive part in the circulation system is pumped to the outlet A, the middle adsorptive part to the outlet B, and the strong The positional relationship between the suction port C of the absorbent part and the supply port F of the original solution is the specific relationship defined by the present invention, so that the use of a smaller amount of adsorbent can be used to extract the weakly adsorbent components, the intermediate adsorbent components and the Strongly adsorbing ingredients. This example reveals the technical content of obtaining the target antibody with high purity and high efficiency in the middle adsorptive part.

[Example 5] Multi-column, gradient, and simulated moving layer method ＜Pipe string＞ Diameter 10mm × length 80mm, 5 pieces. ＜Original solution＞ The concentration of NaCl in the stock solution is 2.46g/L. ＜Elute solution＞ Use the following eluent.

[Table 26] Table 26 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.46 D3 0.00 D4 1.98

Use the combination of the sub-steps shown in Figure 13 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, execute 5 sets of them as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 28] Table 28 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Example 6] Multi-tubular column, gradient, and simulated moving layer method <Tube column> Diameter 10mm×Length 80mm, 5 pieces. <The stock solution> The concentration of NaCl in the stock solution is 2.55g/L. <Eluent> The following eluent was used. [Table 29] Table 29 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.55 D3 0.00 D4 2.03

<Operating conditions> Use the combination of the sub-steps shown in Figure 15 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, execute 5 sets of them as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 31] Table 31 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle adsorptive part antibody 98.0 Strong adsorption part Aggregate 98.0

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

[Example 7] Multi-tubular column, gradient, simulated moving layer method <Tube column> Diameter 10mm×Length 80mm, 5 pieces. <The stock solution> The concentration of NaCl in the stock solution is 2.55g/L. <Eluent> The following eluent was used. [Table 32] Table 32 Type of dissolving liquid NaCl concentration of eluate (g/L) D1 17.53 D2 2.55 D3 0.00 D4 2.03

<Operating conditions> Use the combination of the sub-steps shown in Figure 17 to form step (A). Take this step (A) and the subsequent step (B) as a set of steps, execute 5 sets of them as 1 cycle, and implement 10 cycles. The supply flow rates of the eluent (D1 to D4) and the stock solution (F) in each step (A) are as follows.

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

＜Results＞ -Recovery rate- The recovery rates of fragments 1 and 2 of the weakly adsorbing portion, the recovery rate of antibodies in the middle adsorbing portion, and the recovery rate of aggregates of the strongly adsorbing portion are shown in the following table.

[Table 34] Table 34 section Recycled components Recovery rate(%) Weak adsorption part Fragment 1 Over 99 Fragment 2 98.0 Middle 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 and its implementation are described together, unless otherwise specified, the present invention is not limited by a certain detail in the description, and should not violate the invention disclosed in the appended claims. Explain it in the context of spirit and scope.

In this case, priority is claimed based on the patent application No. 2018-215950 filed in Japan on November 16, 2018 and the patent application No. 2019-088523 filed in Japan on May 8, 2019, which is referred to here. And other cases and take its content as part of the description of this manual.

111: Piping 1~7: The number of the unit packing tower 2a, 2b, 2c, 2d: The weakly adsorbing part is withdrawn from the pipeline 2J: Weakly adsorbing part of the junction pipe 3a, 3b, 3c, 3d: the middle adsorptive part is withdrawn from the pipeline 3J: Mid-adsorption part confluence pipe 4a, 4b, 4c, 4d: strong adsorption part withdrawable from the pipeline 4J: Partial junction pipe with strong adsorption 116: stock tank 117: Original Solution 8a, 8b, 8c, 8d: eluent tank 9a, 9b, 9c, 9d: eluent 10a, 10b, 10c, 10d: unit packed tower (pipe string) 11a, 11b, 11c, 11d: dope supply branch pipeline 11: Stock solution supply line 12, 13, 14, 15: eluent supply line 12a, 12b, 12c, 12d: eluent supply branch pipeline 13a, 13b, 13c, 13d: eluent supply branch pipeline 14a, 14b, 14c, 14d: eluent supply branch pipeline 15a, 15b, 15c, 15d: eluent supply branch pipeline 100: Circulatory system (A): Steps (A1-1), (A2-1), (A3-1), (A1-2), (A2-2), (A3-2), (A1-3), (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-step A1, A2, A3, A4: Weak absorption part extraction valve Ab: adsorbent A: The weakly absorbent part is pumped out B1, B2, B3, B4: Extraction valve for the middle adsorptive part B: Pumping outlet for the medium adsorptive part C1, C2, C3, C4: strong adsorption part extraction valve C: Part of the strong adsorption outlet D1, D2, D3, D4, D5: eluent supply port E1a, E2a, E3a, E4a: eluent supply valve E1b, E2b, E3b, E4b: eluent supply valve E1c, E2c, E3c, E4c: eluent supply valve E1d, E2d, E3d, E4d: eluent supply valve F1, F2, F3, F4: stock liquid supply valve F: Stock solution supply port P1: circulation pump P2: Dosing liquid supply pump P3, P4, P5, P6: eluent supply pump R1, R2, R3, R4: blocking valve T1, T2, T3, T4: check valve

[Figure 1] is a system diagram showing an example of the chromatographic separation system of the simulated moving layer method of the present invention. [Figure 2] is a flowchart of each sub-step constituting step (A) in an embodiment of the chromatographic separation method of the simulated moving layer method of the present invention. [Figure 3] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Figure 2 and implementing step (B). [Fig. 4] It is a flowchart of each sub-step constituting step (A) in another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Figure 5] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Figure 4 and implementing step (B). [Fig. 6] It is a flowchart of each sub-step that constitutes step (A) in still another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Fig. 7] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Fig. 6 and implementing step (B). [Fig. 8] It is a flowchart of each sub-step constituting step (A) in still another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Fig. 9] It is a flowchart of each sub-step of step (A) after step (B) after finishing step (A) shown in Fig. 8. [Fig. 10] A flowchart showing the operation procedure of the single string and step gradient in Comparative Examples 1 and 2. [FIG. 11] A flowchart showing the operation procedure of chromatographic separation in a simulated moving layer method in Comparative Example 3. [FIG. 12] It is a flowchart showing the operation procedure of chromatographic separation in a simulated moving layer method in Comparative Example 4. [FIG. 13] It is a flowchart of each sub-step constituting step (A) in still another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Fig. 14] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Fig. 13 and implementing step (B). [FIG. 15] It is a flowchart of each sub-step of step (A) in still another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Figure 16] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Figure 15 and implementing step (B). [FIG. 17] It is a flowchart of each sub-step of step (A) in still another embodiment of the chromatographic separation method of simulated moving layer mode of the present invention. [Fig. 18] It is a flowchart of each sub-step of step (A) after finishing step (A) shown in Fig. 17 and implementing step (B).

111: Piping

2a, 2b, 2c, 2d: The weakly adsorbing part is withdrawn from the pipeline

2J: Weakly adsorbing part of the junction pipe

3a, 3b, 3c, 3d: the middle adsorptive part is withdrawn from the pipeline

3J: Mid-adsorption part confluence pipe

4a, 4b, 4c, 4d: strong adsorption part withdrawable from the pipeline

4J: Partial junction pipe with strong adsorption

116: stock tank

117: Original Solution

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

9a, 9b, 9c, 9d: eluent

10a, 10b, 10c, 10d: unit packed tower (pipe string)

11a, 11b, 11c, 11d: dope supply branch pipeline

11: Stock solution supply line

12, 13, 14, 15: eluent supply line

12a, 12b, 12c, 12d: eluent supply branch pipeline

13a, 13b, 13c, 13d: eluent supply branch pipeline

14a, 14b, 14c, 14d: eluent supply branch pipeline

15a, 15b, 15c, 15d: eluent supply branch pipeline

100: Circulatory system

A1, A2, A3, A4: Weak absorption part extraction valve

Ab: adsorbent

B1, B2, B3, B4: Extraction valve for the middle adsorptive part

C1, C2, C3, C4: strong adsorption part extraction valve

E1a, E2a, E3a, E4a: eluent supply valve

E1b, E2b, E3b, E4b: eluent supply valve

E1c, E2c, E3c, E4c: eluent supply valve

E1d, E2d, E3d, E4d: eluent supply valve

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

P1: circulation pump

P2: Dosing liquid supply pump

P3, P4, P5, P6: eluent supply pump

R1, R2, R3, R4: blocking valve

T1, T2, T3, T4: check valve

Claims (13)

A method for chromatographic separation in a simulated moving layer method, which includes a circulation system in which a plurality of unit packed towers filled with adsorbents are connected in series and endlessly through pipes, and the raw liquid contained in the original solution is weaker than the adsorbent. Two or more eluents are used to separate the adsorptive components, strong adsorptive components, and intermediate adsorptive components in the middle of the two components; the characteristics of this simulated moving-layer chromatography separation method are: The piping of the circulation system is provided with a raw liquid supply port F, two or more eluent supply ports D corresponding to each eluent of two or more types, and extraction of the weakly adsorbable part containing the weakly adsorbable component Port A, the suction port B of the middle adsorptive portion containing the middle adsorptive component, and the suction port C of the strongly adsorptive portion containing the strongly adsorptive component; And the positions of the stock solution supply port F, the pumping port A, the pumping port B, and the pumping port C are set to the following (a) to (c): (A) The extraction port B is provided on the downstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (B) The extraction port C is provided in the piping having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (C) The extraction port A is provided in the piping with the extraction port B, or the extraction port A is provided on the downstream side of the extraction port B sandwiching at least one unit packing tower; The chromatographic separation method includes a simulated moving layer chromatographic separation method including steps (A) and (B), which are repeated in sequence: [Step (A)] The stock solution is supplied from the stock solution supply port F at the same time or separately, two or more types of eluent are supplied from the two or more eluent supply ports D, and the weak adsorption is drawn from the extraction port A simultaneously or separately Part, the step of extracting the medium adsorptive part from the extraction port B and extracting the strong adsorptive part from the extraction port C [Step (B)] After the step (A) is completed, the raw liquid supply port F, the eluent supply port D, the pumping port A, the pumping port B, and the pumping port C are made to move downstream while maintaining their relative positional relationship Steps to move sideways. Such as the chromatographic separation method of simulated moving layer mode of claim 1, wherein: This step (A) consists of multiple 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. Such as the chromatographic separation method of simulated moving layer mode of claim 1 or 2, wherein, The extraction port C is provided on the downstream side of the eluent supply port D1 that supplies the eluent d1 with the strongest desorption force among the two or more eluents; At least one unit packed tower is arranged between the eluent supply port D1 and the suction port C; In this step (A), during the supply of the eluent d1, from the extraction port C, a strong adsorbent portion of the same amount as the supply amount of the eluent d1 is extracted. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 3, wherein: The suction port B is provided on the downstream side of the eluent supply port D2 that supplies the eluent d2 with the second strongest desorption force among the two or more eluents; At least one unit packed tower is arranged from the eluent supply port D2 to the suction port B; In this step (A), during the supply of the eluent d2, a time zone in which the same amount of the neutral adsorptive portion as the supply amount of the eluent d2 is extracted from the extraction port B is provided. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 4, wherein: Use 4 to 6 types of eluents with different desorption powers. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 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 an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. , In addition, use the two or more eluents, and perform the following sub-steps (A1-1), (A2-1) and (A3-1) in this step (A): ＜Substep (A1-1)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, and uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of section 3 The end is used as the raw liquid supply port F, the raw liquid is supplied from the raw liquid supply port F, the downstream end of the section 4 is used as the suction port A, and the weakly adsorbent portion is extracted from the suction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in section 2 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent flowing in sections 3 and 4 weaker than that of the eluent flowing in section 2; ＜Sub-step (A2-1)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The upstream end of the section 3 serves as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorbing portion is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in section 2 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent flowing in sections 3 and 4 weaker than that of the eluent flowing in section 2; <Sub-step (A3-1)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 3 is used as the suction port B, and the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 4 is used as the eluent supply port D-IV, from the eluent supply port D-IV The eluent d-IV is supplied, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 4 is weaker than that of the eluent flowing in sections 2 and 3. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 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 an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. , In addition, use these two or more eluents, and perform the following sub-steps (A1-2), (A2-2) and (A3-2) in this step (A): ＜Substep (A1-2)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The raw liquid supply port F supplies raw liquid, and the downstream end of the section 4 is used as the suction port A, and the weakly adsorbing part is drawn from the suction port A, thereby, Make the desorption force of the eluent flowing in the zones 1 and 2 the strongest, Make the desorption force of the eluent circulating in sections 3 and 4 weaker than that of the eluent circulating in sections 1 and 2; <Sub-step (A2-2)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, and uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of section 3 The end is used as the eluent supply port D-III, the eluent d-III is supplied from the eluent supply port D-III, and the weakly adsorbing portion is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in section 2 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent flowing in sections 3 and 4 weaker than that of the eluent flowing in section 2; <Substep (A3-2)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-2) For the eluent d-II, the downstream end of section 3 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of section 4 is used as the eluent supply port D-IV from The eluent supply port D-IV supplies the eluent d-IV, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 4 is weaker than that of the eluent flowing in sections 2 and 3. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 5, wherein: The circulation system has 5 or more unit packed towers, and each zone has at least one unit packed tower, and the circulation system is divided into 5 zones 1 to 5 connected in an annular shape from the upstream side to the downstream side. , In addition, use these two or more eluents, and perform the following sub-steps (A1-3), (A2-3) and (A3-3) in this step (A): <Sub-step (A1-3)> The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 serves as the eluent supply port D-III, the eluent supply port D-III supplies eluent d-III, and the downstream end of section 5 As the extraction port A, the weakly adsorbing part is extracted from the extraction port A, thereby, Make the desorption force of the eluent flowing in the zones 1 and 2 the strongest, Make the desorption force of the eluent flowing in zone 3 the same as or weaker than the desorption force of the eluent flowing in zones 1 and 2, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; ＜Substep (A2－3)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II, and supplies it from the eluent The port D-III supplies the eluent d-III, and the weakly adsorbable part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent circulating in sections 4 and 5 weaker than that of the eluent circulating in sections 2 and 3; ＜Substep (A3－3)＞ The eluent d-I is supplied from the eluent supply port D-1, the strongly adsorbing part is extracted from the suction port C, and supplied from the eluent supply port D-II in the substep (A2-3) For the eluent d-II, the downstream end of the section 4 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV. The eluent supply port D-IV supplies the eluent d-IV, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2, 3, and 4 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 5 is weaker than that of the eluent flowing in sections 2, 3, and 4. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 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 an annular shape from the upstream side to the downstream side so that each section has at least one unit packed tower. , In addition, use the two or more eluents, and perform the following sub-steps (A1-4), (A2-4) and (A3-4) in this step (A): ＜Substep (A1-4)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the upstream end of the section 3 is used as the raw solution supply port F. The stock solution supply port F supplies stock solution, the upstream end of section 4 serves as the eluent supply port D-III, the eluent supply port D-III supplies eluent d-III, and the downstream end of section 5 As the extraction port A, the weakly adsorbing part is extracted from the extraction port A, thereby, Make the desorption force of the eluent flowing in the zones 1 and 2 the strongest, Make the desorption force of the eluent flowing in zone 3 the same as or weaker than the desorption force of the eluent flowing in zones 1 and 2, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; ＜Substep (A2－4)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II to the upstream side of the section 4 The end is used as the eluent supply port D-IV, the eluent d-IV is supplied from the eluent supply port D-IV, and the weakly adsorbing part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent circulating in sections 4 and 5 weaker than that of the eluent circulating in sections 2 and 3; <Substep (A3-4)> The eluent d-I is supplied from the eluent supply port D-1, the strongly adsorbing part is extracted from the extraction port C, and supplied from the eluent supply port D-II in the substep (A2-4) For the eluent d-II, the downstream end of the section 4 is used as the suction port B, the middle adsorptive part is drawn from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-V, from The eluent supply port DV supplies the eluent d-V, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2, 3, and 4 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 5 is weaker than that of the eluent flowing in sections 2, 3, and 4. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 5, wherein: The circulation system has 5 or more unit packed towers, and each zone has at least one unit packed tower, and the circulation system is divided into 5 zones 1 to 5 connected in an annular shape from the upstream side to the downstream side. In addition, use the two or more eluents, and perform the following sub-steps (A1-5), (A2-5) and (A3-5) in this step (A): <Substep (A1-5)> The upstream end of the section 3 is used as the stock solution supply port F, the stock solution is supplied from the stock solution supply port F, and the upstream end of the section 4 is used as the eluent supply port D-III, from the eluent supply port D-III The eluent d-III is supplied, the downstream end of the section 5 is used as the suction port A, and the weakly adsorptive part is extracted from the suction port A, thereby, Make the desorption force of the eluent circulating in zone 3 the strongest, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; ＜Substep (A2－5)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of section 2 as the eluent supply port D-II, and supplies the eluent d-II from the eluent supply port D-II, and supplies it from the eluent The port D-III supplies the eluent d-III, and the weakly adsorbable part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent circulating in sections 4 and 5 weaker than that of the eluent circulating in sections 2 and 3; ＜Substep (A3－5)＞ The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 4 serves as the suction port B, and the middle adsorptive part is extracted from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV, and from the eluent supply port D-IV The eluent d-IV is supplied, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2, 3, and 4 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 5 is weaker than that of the eluent flowing in sections 2, 3, and 4. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 5, wherein: The circulation system has 5 or more unit packed towers, and each zone has at least one unit packed tower, and the circulation system is divided into 5 zones 1 to 5 connected in an annular shape from the upstream side to the downstream side. In addition, use the two or more eluents, and perform the following sub-steps (A1-6), (A2-6) and (A3-6) in this step (A): ＜Substep (A1－6)＞ The upstream end of section 1 is used as the eluent supply port D-II, the eluent d-II is supplied from the eluent supply port D-II, and the downstream end of section 3 is used as the suction port B. The suction port B draws out the middle adsorptive part, and uses the upstream end of section 4 as the eluent supply port D-IV, and supplies the eluent d-IV from the eluent supply port D-IV to the downstream side of section 5 The end serves as the extraction port A, and the weakly adsorbing part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in the zones 1, 2 and 3 strongest, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in sections 1, 2 and 3; <Substep (A2-6)> The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 3 as the stock solution supply port F, supplies the stock solution from the stock solution supply port F, and uses the upstream end of the section 4 as the eluent supply port D-III, from The eluent supply port D-III supplies the eluent d-III, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in zone 3 weaker than that of the eluent flowing in zone 1, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; <Substep (A3-6)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than the desorption force of the eluent flowing in sections 2 and 3. Such as the chromatographic separation method in simulated moving layer mode according to any one of claims 1 to 5, wherein: The circulation system has 5 or more unit packed towers, and each zone has at least one unit packed tower, and the circulation system is divided into 5 zones 1 to 5 connected in an annular shape from the upstream side to the downstream side. In addition, use the two or more eluents, and perform the following sub-steps (A1-7), (A2-7) and (A3-7) in this step (A): ＜Substep (A1-7)＞ The upstream end of section 1 is used as the eluent supply port D-1, the eluent d-I is supplied from the eluent supply port D-I, and the downstream end of section 1 is used as the suction port C. The suction port C draws out the strongly adsorbing part, uses the upstream end of the section 3 as the stock solution supply port F, supplies the stock solution from the stock solution supply port F, and uses the upstream end of the section 4 as the eluent supply port D-III, from The eluent supply port D-III supplies the eluent d-III, the downstream end of the section 5 is used as the suction port A, and the weakly adsorbing portion is extracted from the suction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in zone 3 weaker than that of the eluent flowing in zone 1, Make the desorption force of the eluent flowing in sections 4 and 5 weaker than that of the eluent flowing in section 3; <Substep (A2-7)> The eluent d-I is supplied from the eluent supply port D-1, and the strongly adsorbing portion is extracted from the suction port C. The upstream end of the zone 2 is used as the eluent supply port D-II, and the eluent The eluent supply port D-II supplies the eluent d-II, the eluent liquid d-III is supplied from the eluent supply port D-III, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2 and 3 weaker than that of the eluent flowing in section 1, Make the desorption force of the eluent circulating in sections 4 and 5 weaker than that of the eluent circulating in sections 2 and 3; <Substep (A3-7)> The eluent d-I is supplied from the eluent supply port D-I, the strongly adsorbent portion is extracted from the suction port C, and the eluent d-II is supplied from the eluent supply port D-II to The downstream end of the section 4 serves as the suction port B, and the middle adsorptive part is extracted from the suction port B, and the upstream end of the section 5 is used as the eluent supply port D-IV, and from the eluent supply port D-IV The eluent d-IV is supplied, and the weakly adsorptive part is extracted from the extraction port A, thereby, Make the desorption force of the eluent circulating in zone 1 the strongest, Make the desorption force of the eluent flowing in sections 2, 3, and 4 weaker than that of the eluent flowing in section 1, The desorption force of the eluent flowing in section 5 is weaker than that of the eluent flowing in sections 2, 3, and 4. A simulated moving-layer chromatography separation system, which uses a plurality of unit packed towers filled with adsorbents in a circulatory system connected in series and endlessly through piping to weakly adsorb the raw liquid contained in the adsorbent against the adsorbent Two or more eluents are used to separate the chromatographic separation system with two or more eluents; the characteristics of this simulated moving-layer chromatography separation system are: The piping of the circulation system is provided with a raw liquid supply port F, two or more eluent supply ports D corresponding to each eluent of two or more types, and extraction of the weakly adsorbable part containing the weakly adsorbable component Port A, the suction port B of the middle adsorptive portion containing the middle adsorptive component, and the suction port C of the strongly adsorptive portion containing the strongly adsorptive component; And the positions of the stock solution supply port F, the pumping port A, the pumping port B, and the pumping port C are set to the following (a) to (c): (A) The extraction port B is provided on the downstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (B) The extraction port C is provided in the piping having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F sandwiching at least one unit packing tower; (C) The extraction port A is provided in the piping with the extraction port B, or the extraction port A is provided on the downstream side of the extraction port B sandwiching at least one unit packing tower; The chromatographic separation system is a simulated moving layer chromatographic separation system with a mechanism that sequentially repeats the following steps (A) and (B): [Step (A)] The stock solution is supplied from the stock solution supply port F at the same time or separately, two or more types of eluent are supplied from the two or more eluent supply ports D, and the weak adsorption is drawn from the extraction port A simultaneously or separately Part, the step of extracting the medium adsorptive part from the extraction port B and extracting the strong adsorptive part from the extraction port C [Step (B)] After the step (A) is completed, the raw liquid supply port F, the eluent supply port D, the pumping port A, the pumping port B, and the pumping port C are made to move downstream while maintaining their relative positional relationship Steps to move sideways.

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