Continuous chromatography system based on tangential flow chromatography columns and methods of use thereof

Abstract

The application provides a continuous chromatography system based on tangential flow chromatography columns and a use method thereof, which comprises a sample loading pipeline, an elution pipeline and a plurality of tangential flow chromatography columns; a sample loading pump is arranged on the sample loading pipeline; the outlet end of the sample loading pipeline is connected with the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; an elution pump is arranged on the elution pipeline; the outlet end of the elution pipeline is connected with the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; the inlet and outlet end pipelines at the top of each tangential flow chromatography column are connected with a first collecting pipeline and a second collecting pipeline through valves respectively; the backflow port pipeline of the tangential flow chromatography column is connected with the sample loading pipeline through a filtering module or a liquid storage tank; in use, one is in a loading mode and the other is in an elution mode; the application has the advantages that there is almost no residual substance in the flow path and the circulation loop, and the circulation loop does not have to be stopped and restarted for many times.

Description

TECHNICAL FIELD

[0001] The present application relates to the downstream separation and purification of biopharmaceuticals, in particular to a continuous chromatography system based on tangential flow chromatography columns and a method of using the same. BACKGROUND

[0002] The chromatography columns used in the downstream of biopharmaceuticals are all structured so that the sample to be processed enters from one end of the column and is collected from the other end, so they cannot handle particulate precipitates and cannot backflush the distribution screen plates, and cannot achieve the ability to perform high-performance chromatography and particle processing simultaneously. In the prior art, for example, Patent No. 202120067489.X, a hydraulic drive type dynamic chromatography device, when the separation liquid needs to be added, the liquid is injected from the liquid inlet and collected from the liquid outlet; or uses, for example, Patent No. 201820028656.8, a dynamic beam type full-automatic chromatography column, the bottom uses a structure of a screen plate, a flow distribution plate, and a base, the center of the screen plate is provided with a lower nozzle, when the separation liquid needs to be added, the liquid enters from the chromatography port of the upper nozzle and is collected from the chromatography port of the lower nozzle, or enters from the chromatography port of the lower nozzle and is collected from the chromatography port of the upper nozzle. The disadvantages are: 1. Cannot realize column head upper flushing or elution; 2. Does not support reverse flushing of the filter plate (screen plate), which is prone to blockage; 3. Cannot simultaneously achieve high linear flow rate and overload loading, and has low dynamic capacity.

[0003] The present applicant proposes a tangential flow chromatography column, Patent No. 202220697921.8, Patent Name: A radial and tangential flow bottom for chromatography columns and a chromatography system using the same. The tangential flow structure is adopted to realize chromatography loading while performing tangential flow filtration, thereby realizing high linear flow rate and overload loading, and achieving extremely high dynamic capacity. Therefore, in order to meet the production needs, a continuous flow chromatography system using the tangential flow chromatography column is needed. SUMMARY

[0004] In order to achieve the above-mentioned purpose, the present application provides a continuous chromatography system based on tangential flow chromatography columns and a method of using the same, which can realize that part of the chromatography columns are in loading mode and part are in elution mode, so that the sample can be continuously loaded in the tangential flow chromatography column, and there is no residual in the flow path and the circulation loop.

[0005] In order to achieve the above-mentioned purpose, the technical solution adopted by the present application is:

[0006] The first aspect of the present application provides a continuous chromatography system based on tangential flow chromatography column, comprising a sample loading pipeline, an elution pipeline and a plurality of tangential flow chromatography columns; the sample loading pipeline is provided with a sample loading pump, and the outlet end of the sample loading pipeline is connected to the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; the elution pipeline is provided with an elution pump, and the outlet end of the elution pipeline is connected to the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; the inlet and outlet end pipelines at the top of each tangential flow chromatography column are connected to a first collection pipeline and a second collection pipeline through valves respectively; and the backflow port pipelines of the tangential flow chromatography columns are connected to the sample loading pipeline through a filter module or a liquid storage tank.

[0007] In some embodiments, the tangential flow chromatography columns are two, one in loading mode and the other in elution mode.

[0008] In some embodiments, the outlet end pipelines of the sample loading pipeline and the elution pipeline are respectively connected to a first valve array; the first valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array correspond to the outlet end pipelines of the sample loading pipeline and the elution pipeline respectively; and the other pair of opposite sides of the rectangular valve array correspond to the inlet and outlet end pipelines at the bottom of the two chromatography columns respectively.

[0009] In some embodiments, the top inlet and outlet end pipelines of the two chromatography columns are respectively connected to a second valve array; the first valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array correspond to the top inlet and outlet end pipelines of the two chromatography columns respectively; and the other pair of opposite sides of the rectangular valve array correspond to the inlet ends of the first collection pipeline and the second collection pipeline.

[0010] In some embodiments, the filter module comprises a plurality of filters; one end of each of the filters is connected to the backflow port pipelines of the two tangential flow chromatography columns through valves respectively; and the other end of each of the filters is connected to the sample loading pipeline.

[0011] In some embodiments, the filters are two.

[0012] In some embodiments, the backflow end pipelines of the two tangential flow chromatography columns are connected to the two filters through a third valve array; the third valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array correspond to the inlet and outlet end pipelines of one end of the two filters respectively; and the other pair of opposite sides of the rectangular valve array correspond to the backflow end pipelines and a waste discharge pipeline respectively; and the other end of the two filters is connected to the sample loading pipeline through a fourth valve array; the fourth valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array correspond to the inlet and outlet end pipelines of the other end of the two filters respectively; and the other pair of opposite sides of the rectangular valve array correspond to a liquid inlet pipeline and the sample loading pipeline.

[0013] In some embodiments, a backflow port pipeline of the tangential flow chromatography column is provided with a backflow pump.

[0014] In some embodiments, an outlet pipeline of the filter module or the liquid storage tank is connected to the sample loading pipeline through a mixer.

[0015] In some embodiments, the outlet end of the elution pipeline is connected to the liquid inlet pipeline at the top of each tangential flow chromatography column through a valve.

[0016] The second aspect of the present application provides a method for using a tangential flow chromatography column-based continuous chromatography system, which comprises the following steps: sample loading and elution; sample loading: the sample to be treated is introduced into the bottom inlet and outlet of a part of the tangential flow chromatography columns through the sample loading pipeline, part of the solution is backflowed to the sample loading pipeline through the backflow port of the tangential flow chromatography column to form a circulation after passing through the filter module or the liquid storage tank, and the waste solution is collected in the first collection pipeline after passing through the tangential flow chromatography column; elution: the eluent is introduced into the bottom inlet and outlet of the remaining part of the tangential flow chromatography columns through the elution pipeline, and the solution is collected in the second collection pipeline after passing through the tangential flow chromatography column; then, the system is alternately operated, with one part in the loading mode and the other part in the elution mode.

[0017] In some embodiments, two tangential flow chromatography columns are used, one in the loading mode and the other in the elution mode.

[0018] In some embodiments, the outlet end of the elution pipeline is connected to the liquid inlet pipeline at the top of each tangential flow chromatography column through a valve.

[0019] In some embodiments, the method for using the tangential flow chromatography column-based continuous chromatography system further comprises post-loading flushing: the buffer is introduced into the top inlet and outlet of the tangential flow chromatography column after sample loading through the elution pipeline, and the solution is collected in the collection pipeline connected to the backflow port pipeline after passing through the tangential flow chromatography column.

[0020] In some embodiments, the method for using the tangential flow chromatography column-based continuous chromatography system further comprises post-elution regeneration: the cleaning agent is introduced into the top inlet and outlet of the tangential flow chromatography column after elution through the elution pipeline, and the solution is collected in the collection pipeline connected to the backflow port pipeline after passing through the tangential flow chromatography column.

[0021] The present application has the following advantages:

[0022] 1. In the present invention two tangential flow chromatography columns are used, which are operated alternately: one in loading mode and the other in elution mode. By doing this, the feed can be continuously loaded into the RTF columns, with almost no leftovers in the flow path and the circulation loop, which does not have to be stopped and restarted multiple times.

[0023] 2. The return end of the tangential flow uses a circulation loop, which is driven while loading the tangential flow chromatography columns.

[0024] 3. With the filtration module it removes particulate matter or contaminants from the feed and keeps the particulate matter concentration constant (lower than the feed concentration). BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 Figure 1 shows the schematic structure of the present invention. DETAILED DESCRIPTION

[0026] The present invention will be further described with reference to the drawings.

[0027] The technical content of the present invention is described below through specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the disclosure of the specification. The present invention can also be implemented or applied through other different embodiments. Those skilled in the art can make various modifications and changes without departing from the spirit of the present invention.

[0028] Before describing the specific embodiments of the present disclosure in detail, first, some terms used in the present disclosure are explained.

[0029] Unless otherwise defined in the following, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Reference herein to technical terms used herein is intended to refer to the technical terms as commonly understood by those skilled in the art, including variations or substitutions of the technology that are obvious to those skilled in the art or equivalent replacements of the technology. Although the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention. When a term appears herein, it is intended to refer to its corresponding term. All patents, published patent applications, and publications cited herein are incorporated by reference herein.

[0030] The terms "connected," "connected to," "coupled," "coupled to," or "coupled with" or similar terms as used herein are not limited to direct connections, but also include indirect connections.

[0031] A "sample" as described herein is a biomolecule, including proteins, nucleic acids, lipids, carbohydrates, small nucleotides, amino acids, and derivatives thereof.

[0032] The term "on-line monitoring" or "real-time monitoring" as used herein refers to detecting certain parameters or properties of the buffer, reaction fluid, fluid flowing out of the flow reactor, such as pH value, pressure, flow rate, conductivity, etc. in real time during the use of the chromatography system. Unlike off-line detection or analysis, on-line monitoring or real-time monitoring can provide real-time feedback of the detection results.

[0033] The positional relationship words "upper", "lower", "left", "right", "front", "back", etc. as referred to herein are determined according to the layout direction of the drawings of the specification, which are only used to represent relative positional relationship, and when the absolute position of the described object is changed, the relative positional relationship may also be changed accordingly.

[0034] The RTF chromatography column (tangential flow chromatography column) as referred to herein is a chromatography column independently researched and developed by the company and has a patent, with a patent number of 202220697921.8; having a radial and tangential flow structure, which can be loaded through tangential flow at the bottom of the column, and the upper part of the column head can be simultaneously washed and eluted, and the mode of circulating loading and washing with equilibrium liquid is used, so as to realize high linear flow rate and overload loading, and realize extremely high dynamic capacity.

[0035] Referring to Figure 1 As shown in the figure, a continuous chromatography system based on a tangential flow chromatography column includes a loading pipeline 1 and an elution pipeline 2. The inlet end of the loading pipeline 1 is connected to a first tank 3 through a valve AV1, and the first tank 3 can be used to store samples to be processed. The inlet end of the loading pipeline 1 is connected to a second tank 4 through a valve AV4. A loading pump 5 and a first flow indicating transmitter 6 are arranged on the loading pipeline 1. The outlet end of the loading pipeline 1 is connected to a first valve array 7, which is a rectangular valve array composed of a valve AV10, a valve AV11, a valve AV12, and a valve AV13 connected end to end. The loading pipeline 1 is connected between the valve AV10 and the valve AV11. The inlet end of the elution pipeline 2 is connected to the second tank 4 through a valve AV5, to a third tank 8 through a valve AV2, and to a fourth tank 9 through a valve AV3. An elution pump 10, a second flow indicating transmitter 11, and a bubble trap 33 are arranged on the elution pipeline 2. The outlet end of the elution pipeline 2 is connected between the valve AV12 and the valve AV13 through a pipeline, and the pipeline is provided with a valve AV14. The valve AV10 and the valve AV12 are connected through a pipeline to the bottom inlet and outlet ends of a first tangential flow chromatography column 12; and the valve AV11 and the valve AV13 are connected through a pipeline to the bottom outlet end of a second tangential flow chromatography column 13.

[0036] The top inlet and outlet lines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 are connected to the second valve array 14, which is a rectangular valve array composed of valve AV17, valve AV18, valve AV19 and valve AV20. The top inlet and outlet lines of the first tangential flow chromatography column 12 are connected between valve AV19 and valve AV20, and the top inlet and outlet lines of the second tangential flow chromatography column 13 are connected between valve AV17 and valve AV18. Valve AV17 and valve AV19 are connected to the first collection pipeline 15; valve AV18 and valve AV20 are connected to the second collection pipeline 16. The first collection pipeline 15 is provided with sensors for real-time monitoring of data: first UV sensor 17, first conductivity transmitter 18, first pH sensor 19. The second collection pipeline 16 is provided with sensors for real-time monitoring of data: second UV sensor 20, second conductivity transmitter 21, second pH sensor 22.

[0037] The return port line of the first tangential flow chromatography column 12 is connected to the inlet end of the connection pipeline 23 through valve AV6, and the return port line of the second tangential flow chromatography column 13 is connected to the connection pipeline 23 through valve AV7. The connection pipeline 23 is provided with a circulating pump 24 and a third flow indication transmitter 25. The outlet end of the connection pipeline 23 is connected to the third valve array 26, which is a rectangular valve array composed of pressure control valve PCV1, pressure control valve PCV2, valve AV26 and valve AV27. The connection pipeline 23 is connected between pressure control valve PCV1 and pressure control valve PCV2; the first inlet and outlet lines of the first filter 27 are connected between pressure control valve PCV1 and valve AV26; the first inlet and outlet lines of the second filter 28 are connected between pressure control valve PCV2 and valve AV27; the waste discharge line 29 is connected between valve AV26 and valve AV27. The second inlet and outlet lines of the first filter 27 and the second filter 28 are connected to the fourth valve array 30, which is a rectangular valve array composed of valve AV22, valve AV23, valve AV24 and valve AV25. The second inlet and outlet lines of the first filter 27 are connected between valve AV22 and valve AV23; the second inlet and outlet lines of the second filter 28 are connected between valve AV24 and valve AV25; the liquid inlet pipeline 31 is connected between valve AV22 and valve AV24; the sample loading pipeline 1 is connected between valve AV23 and valve AV25 through a pipeline; valve AV21 is arranged on the left side of the connection point of the pipeline and the sample loading pipeline 1.

[0038] The outlet end of the elution pipeline 2 is connected to the top inlet and outlet lines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 through pipelines, respectively, and valve AV16 and valve AV15 are arranged on the pipelines, respectively.

[0039] The bottom backflow port of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 is connected to a solution collection pipeline through valves AV18 and AV19 respectively.

[0040] A mixer 32 is arranged between the sample loading pipeline 1 and the fourth valve array 30.

[0041] A method for using a continuous chromatography system based on tangential flow chromatography columns, the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 can be simultaneously loaded or eluted or one is loaded and the other is eluted.

[0042] 1. When loading simultaneously: the sample to be treated flows out from the first tank 3, enters the sample loading pipeline 1 under the action of the sample loading pump 5, the valves AV10 and AV11 are opened, and the sample to be treated enters the bottom inlet and outlet port pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 respectively, part of the solution enters the connecting pipeline 23 through the bottom backflow port pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, and then enters the filter module through the connecting pipeline 23.

[0043] The filter module uses a first filter 27 and a second filter 28, the two filters can perform filtering work simultaneously or one performs filtering work and the other performs backwashing; or they can perform filtering simultaneously or backwashing simultaneously, for example: the first filter 27 performs filtering and the second filter 28 performs backwashing.

[0044] At this time, the pressure control valve PCV1 and the valve AV23 are opened, the solution flows in through the first inlet and outlet ports of the first filter 27 and flows out through the second inlet and outlet ports of the first filter 27, and finally flows into the sample loading pipeline 1 to form a circulation; the sample to be treated passes through the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, and the waste solution flows out from the top inlet and outlet pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, at this time, the valves AV19 and AV17 are opened, and the waste solution enters the first collection pipeline 15 for collection.

[0045] The second filter 28 performs backwashing: the valves AV24 and AV27 are opened, the WFI solution enters the second inlet and outlet ports of the second filter 28 through the valve AV24, after backwashing, the waste solution enters the waste discharge pipeline 29 through the valve AV27.

[0046] 2. During simultaneous elution: The eluent enters the elution line 2 under the action of the elution pump 10. Valves AV12 and AV13 are opened, and the eluent enters the bottom inlet and outlet pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, respectively. After passing through the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, the solution flows out from the top inlet and outlet pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13. At this time, valves AV20 and AV18 are opened, and the solution enters the second collection pipeline 16 for collection.

[0047] 3. One is loading and the other is eluting: (1) Loading the first tangential flow chromatography column 12 and eluting the second tangential flow chromatography column 13: The sample to be processed flows out from the first tank 3 and enters the loading pipeline 1 under the action of the loading pump 5. The valve AV10 is opened and the sample to be processed enters the bottom inlet and outlet pipelines of the first tangential flow chromatography column 12 respectively. Part of the solution enters the connecting pipeline 23 through the reflux pipeline at the bottom of the first tangential flow chromatography column 12 and enters the filter module through the connecting pipeline 23. The filter module adopts the first filter 27 and the second filter 28. The first filter 27 and the second filter 28 can perform filtration at the same time or one can perform filtration while the other performs backwashing; they can also filter at the same time or perform backwashing at the same time. For example, the first filter 27 is backwashed and the second filter 28 is filtered. At this time, pressure control valve PCV2 and valve AV25 are opened. The solution flows in through the first inlet and outlet ends of the second filter 28 and flows out through the second inlet and outlet ends of the second filter 28, finally flowing into the sample loading line 1 to form a circulation. The sample to be processed passes through the first tangential flow chromatography column 12, and the waste solution flows out from the inlet and outlet lines at the top of the first tangential flow chromatography column 12. At this time, valve AV19 is opened and the waste solution enters the first collection line 15 for collection. The first filter 27 is backwashed: valves AV22 and AV26 are opened. The WFI solution enters through valve AV22 from the second inlet and outlet ends of the first filter 27 for backwashing. After backwashing, the waste solution enters the waste discharge line 29 after passing through valve AV26.

[0048] Simultaneously, the eluent enters the elution line 2 under the action of the elution pump 10. When valve AV13 is opened, the eluent enters the bottom inlet and outlet lines of the second tangential flow chromatography column 13. After passing through the column, the solution flows out from the top inlet and outlet lines of the second tangential flow chromatography column 13. At this time, valve AV18 is opened, and the solution enters the second collection line 16 for collection.

[0049] (2) The first tangential flow chromatography column 12 elution, the second tangential flow chromatography column 13 sample loading: the sample to be processed flows out from the first tank 3, under the action of the sample loading pump 5, enters the sample loading pipeline 1, opens the valve AV11, and the sample to be processed enters the second tangential flow chromatography column 13 bottom inlet and outlet end pipeline, part of the solution enters the connecting pipeline 23 through the reflux end pipeline at the bottom of the second tangential flow chromatography column 13, and enters the filter module through the connecting pipeline 23. The filter module uses a first filter 27 and a second filter 28. The two filters can perform filtration work at the same time or one performs filtration work and the other performs backwashing; or they can be filtered at the same time or backwashed at the same time. For example, the first filter 27 is filtered, and the second filter 28 is backwashed.

[0050] At this time, the pressure control valve PCV1 and the valve AV23 are opened, the solution flows into the first inlet and outlet end of the first filter 27, and the second inlet and outlet end of the first filter 27 flows out, and finally flows into the sample loading pipeline 1 to form a circulation; the sample to be processed passes through the second tangential flow chromatography column 13, and the waste solution flows out from the inlet and outlet pipeline at the top of the second tangential flow chromatography column 13. At this time, the valve AV17 is opened, and the waste solution enters the first collection pipeline 15 for collection.

[0051] The second filter 28 backwashes: the valve AV24 and the valve AV27 are opened, the WFI solution enters the second inlet and outlet end of the second filter 28 from the valve AV24, and after backwashing, the waste solution enters the waste pipeline 29 after passing through the valve AV27.

[0052] At the same time, the eluent enters the elution pipeline 2 under the action of the elution pump 10, the valve AV12 is opened, the eluent enters the first tangential flow chromatography column 12 bottom inlet and outlet end pipeline, passes through the first tangential flow chromatography column 12, and the solution flows out from the inlet and outlet pipeline at the top of the first tangential flow chromatography column 12. At this time, the valve AV20 is opened, and the solution enters the second collection pipeline 16 for collection.

Claims

1. A continuous chromatography system based on a tangential flow chromatography column, characterized in that: It comprises a sample loading pipeline, an elution pipeline and a plurality of tangential flow chromatography columns; the sample loading pipeline is provided with a sample loading pump, and the outlet end of the sample loading pipeline is connected to the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; the elution pipeline is provided with an elution pump, and the outlet end of the elution pipeline is connected to the inlet and outlet end pipelines at the bottom of each tangential flow chromatography column through valves respectively; the inlet and outlet end pipelines at the top of each tangential flow chromatography column are connected to a first collection pipeline and a second collection pipeline through valves respectively; the backflow port pipeline of the tangential flow chromatography column is connected to the sample loading pipeline through a filter module or a liquid storage tank; the outlet end pipelines of the sample loading pipeline and the elution pipeline are connected to a first valve array respectively; the first valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array is connected to the outlet end pipelines of the sample loading pipeline and the elution pipeline respectively; the other pair of opposite sides of the rectangular valve array is connected to the inlet and outlet end pipelines at the bottom of two chromatography columns respectively; the outlet end pipelines of the elution pipeline are connected to the inlet pipelines at the top of a plurality of tangential flow chromatography columns through valves respectively.

2. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The tangential flow chromatography columns adopt two, one in sample loading mode and the other in elution mode.

3. A continuous chromatography system based on a tangential flow chromatography column according to claim 2, characterized in that: The inlet and outlet end pipelines at the top of the two are connected to a second valve array respectively, and the second valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array is connected to the inlet and outlet end pipelines at the top of the two respectively, and the other pair of opposite sides of the rectangular valve array is connected to the inlet end of the first collection pipeline and the second collection pipeline.

4. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The filter module adopts a plurality of filters, and the inlet and outlet at one end of each filter are connected to the backflow port pipelines of two tangential flow chromatography columns through valves respectively, and the other end of the filter is connected to the sample loading pipeline.

5. A continuous chromatography system based on a tangential flow chromatography column according to claim 4, characterized in that: The filter adopts two.

6. A continuous chromatography system based on a tangential flow chromatography column according to claim 5, characterized in that: The backflow end pipelines of the two tangential flow chromatography columns are connected to the two filters through a third valve array, and the third valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array is connected to the inlet and outlet pipelines at one end of the two filters respectively; the other pair of opposite sides of the rectangular valve array is connected to the backflow end pipelines and a waste discharge pipeline respectively; the other end of the two filters is connected to the sample loading pipeline through a fourth valve array, and the fourth valve array is a rectangular valve array formed by four valves connected end to end; one pair of opposite sides of the rectangular valve array is connected to the inlet and outlet pipelines at the other end of the two filters respectively, and the other pair of opposite sides of the rectangular valve array is connected to a liquid inlet pipeline and the sample loading pipeline.

7. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The backflow port pipeline of the tangential flow chromatography column is provided with a backflow pump.

8. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The outlet end pipeline of the filter module or the liquid storage tank is connected to the sample loading pipeline through a mixer.

9. A method of using a continuous chromatography system based on the tangential flow chromatography column according to any one of claims 1 to 8, characterized in that, The steps include: sample loading and elution; sample loading: the sample to be treated enters the bottom inlet and outlet of a part of the tangential flow chromatography column through the sample loading pipeline, part of the solution flows back to the sample loading pipeline through the filter module or the liquid storage tank to form a cycle through the backflow port of the tangential flow chromatography column, after the sample solution to be treated is treated by the tangential flow chromatography column, the waste solution flows into the first collection pipeline from the solution inlet and outlet pipeline at the top of the tangential flow chromatography column for collection; elution: the eluent enters the bottom inlet and outlet of the remaining part of the tangential flow chromatography column that has been loaded through the elution pipeline, after passing through the chromatography column, the solution flows into the second collection pipeline from the solution inlet and outlet pipeline at the top of the tangential flow chromatography column for collection; then alternating operation, a part is in the sample loading mode, and the other part is in the elution mode.

10. A method of using a continuous chromatography system based on a tangential flow chromatography column according to claim 9, characterized in that, The tangential flow chromatography column adopts two, one is in the sample loading mode, and the other is in the elution mode.

11. A method of using a continuous chromatography system based on a tangential flow chromatography column according to claim 10, characterized in that, The outlet end of the elution pipeline is respectively connected to the top liquid inlet pipeline of the two tangential flow chromatography columns through valves.

12. A method of using a continuous chromatography system based on a tangential flow chromatography column according to claim 11, characterized in that, The steps further include post-loading flushing: the buffer enters the top inlet and outlet of the tangential flow chromatography column after sample loading through the elution pipeline, after passing through the chromatography column, the solution flows out from the backflow port of the tangential flow chromatography column, and is collected by the collection pipeline connected to the backflow port pipeline.

13. A method of using a continuous chromatography system based on a tangential flow chromatography column according to claim 11, characterized in that, The steps further include post-elution regeneration: the cleaning agent enters the top inlet and outlet of the tangential flow chromatography column after elution through the elution pipeline, after passing through the chromatography column, the solution flows out from the backflow port of the tangential flow chromatography column, and is collected by the collection pipeline connected to the backflow port pipeline.

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