Chromatographic system based on tangential flow chromatographic column and use method thereof

Abstract

The invention relates to a chromatographic system based on a tangential flow chromatographic column and a use method of the chromatographic system. The chromatographic system comprises a liquid inlet main pipeline and a liquid outlet main pipeline, a liquid inlet pump is arranged on the liquid inlet main pipeline; the outlet end of the liquid inlet main pipeline is respectively connected with inlet and outlet end pipelines at the bottom and the top of the tangential flow chromatographic column through valves; the inlet end of the liquid outlet main pipeline is respectively connected with inlet and outlet pipelines at the bottom and the top of the tangential flow chromatographic column through valves; a backflow end pipeline of the tangential flow chromatographic column is communicated with a liquid inlet main pipeline through a connecting pipeline, and a filtering module is arranged on the connecting pipeline; when in use, the method comprises the steps of balancing, loading a sample, washing after loading the sample and eluting; the device has the advantages that high linear flow velocity and overload sample loading are realized, and extremely high dynamic loading capacity is realized.

Description

Technical Field

[0001] The present invention relates to a chromatography system in the downstream of biomedicine, and in particular to a chromatography system based on a tangential flow chromatography column and a use method thereof. Background Art

[0002] The chromatography columns of the prior art can neither process particulate precipitates, thus resulting in a low chromatographic column flux, nor can they realize backwashing of the distribution plate, and therefore cannot improve the column efficiency and the ability to simultaneously process particles. For example, the patent number is: 02120067489.X, and the utility model name is a hydraulically driven dynamic chromatography device. When separation liquid needs to be added, the liquid is injected from the liquid inlet and flows out from the liquid outlet for collection; or, for example, the patent number is: 201820028656.8, and the utility model name is a moving beam type fully automatic chromatography column, the bottom of which adopts a sieve plate, a diverter plate and a base. A lower nozzle is arranged at the center of the sieve plate. When separation liquid needs to be added, the liquid enters from the chromatography port of the upper nozzle and flows out from the chromatography port of the lower nozzle for collection, or enters from the chromatography port of the lower nozzle and flows out from the chromatography port of the upper nozzle for collection.

[0003] The present applicant provides a tangential flow chromatography column, patent number: 202220697921.8, in which an upper flow channel and a lower flow channel are formed between the confluence plate and the sieve plate, producing a radial and tangential flow structure, so that while realizing tangential flow filtration and chromatographic loading, the sieve plate can be backwashed and is not easily blocked, thereby realizing high linear flow rate and overload loading, and achieving extremely high dynamic loading capacity. At the same time, the use of radial and tangential flow structures can realize tangential flow loading at the bottom of the column, while the upper part of the column head can be flushed and eluted at the same time, cyclic loading and a balanced liquid cleaning mode are used to further improve the dynamic loading capacity. Therefore, for the tangential flow chromatography column provided by the present applicant, it is urgent to provide an application of a chromatography system. Summary of the invention

[0004] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides a chromatography system based on a tangential flow chromatography column and a method of using the same, which can achieve filtration at the reflux end of the tangential flow chromatography column, thereby achieving tangential flow filtration and chromatographic loading at the same time, and the sieve plate can be reversely flushed and is not easy to clog, thereby achieving high linear flow rate and overload loading, and achieving extremely high dynamic loading capacity.

[0005] In order to achieve the above-mentioned purpose, the present invention adopts the following technical solutions:

[0006] The first aspect of the present disclosure provides a chromatography system based on a tangential flow chromatography column, comprising an inlet main pipeline and an outlet main pipeline; the inlet main pipeline is provided with an inlet pump; the outlet end of the inlet main pipeline is connected to the inlet and outlet pipelines at the bottom and top of the tangential flow chromatography column respectively through a valve; the inlet end of the outlet main pipeline is connected to the inlet and outlet pipelines at the bottom and top of the tangential flow chromatography column respectively through a valve; the reflux end pipeline of the tangential flow chromatography column is connected to the inlet main pipeline through a connecting pipeline, and a filtering module is provided on the connecting pipeline.

[0007] Further in some embodiments, the outlet end of the liquid inlet main pipeline is provided with a first valve array, and the first valve array includes a rectangular valve array composed of four valves connected at the ends, and the outlet end of the liquid inlet main pipeline and the inlet end of the liquid outlet main pipeline are respectively connected to a group of opposite sides of the rectangular valve array; the other group of opposite sides of the rectangular valve array are respectively connected to the inlet and outlet pipelines of the corresponding bottom and top of the tangential flow chromatography column.

[0008] In some further embodiments, the filtration module is a filter, the pipelines at the first inlet and outlet ends of the filter are connected to the reflux end pipeline of the tangential flow chromatography column, and the pipelines at the second inlet and outlet ends of the filter are connected to the liquid inlet main pipeline.

[0009] Further in some embodiments, the pipelines at the first inlet and outlet ends of the filter are connected to a waste pipeline, and the pipelines at the second inlet and outlet ends of the filter are connected to a cleaning pipeline.

[0010] In some further embodiments, two filters are used, and the first inlet and outlet ends of the two filters are respectively connected to the reflux end pipeline and the waste pipeline of the tangential flow chromatography column; the second inlet and outlet ends of the two filters are respectively connected to the liquid inlet main pipeline and the cleaning pipeline.

[0011] Further in some embodiments, the first inlet and outlet ends and the second inlet and outlet ends of the two filters are respectively provided with a second valve array and a third valve array, the second valve array is a rectangular valve array composed of four valves connected at the ends, the reflux end pipeline and the waste discharge pipeline of the tangential flow chromatography column are respectively connected to a group of opposite sides of the rectangular valve array, and the other group of opposite sides of the rectangular valve array are respectively connected to the first inlet and outlet ends of the corresponding two filters; the third valve array is a rectangular valve array composed of four valves connected at the ends, one group of opposite sides of the rectangular valve array are respectively connected to the second inlet and outlet ends of the corresponding two filters, and the other group of opposite sides of the rectangular valve array are respectively connected to the corresponding cleaning pipelines and the liquid inlet main pipeline.

[0012] Further in some embodiments, pressure sensors are respectively provided on the connecting pipelines between the reflux end pipeline of the tangential flow chromatography column and the second valve array and between the third valve array and the liquid inlet main pipeline.

[0013] In some further embodiments, a feed end is provided at the top of the tangential flow chromatography column, and the pipelines of the feed end are respectively connected to the top and bottom inlet and outlet pipelines of the filling tank, and valves are respectively provided on the inlet and outlet pipelines; a feed pump is provided on the bottom inlet and outlet pipelines.

[0014] Further in some embodiments, a liquid outlet is provided on one side of the stuffing tank, and the liquid outlet is connected to a liquid inlet main pipeline through a pipeline, and a valve is provided on the pipeline.

[0015] In some further embodiments, a flow indicating transmitter and / or a pressure sensor and / or a conductivity transmitter is provided on the liquid inlet main pipeline.

[0016] In some further embodiments, the liquid outlet main pipeline is provided with a pH sensor and / or a conductivity transmitter and / or a flow indicating transmitter and / or a UV sensor.

[0017] Further in some embodiments, a pressure sensor is provided on the reflux end pipeline of the tangential flow chromatography column.

[0018] The second aspect of the present disclosure provides a method for using a chromatography system based on a tangential flow chromatography column:

[0019] Step 1: Balance, the buffer solution flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the bottom inlet and outlet, enters the liquid outlet main pipeline and is collected;

[0020] Step 2: Loading the sample, the sample solution to be processed flows into the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, part of the sample solution enters the filtration module from the reflux port, and then enters the liquid inlet main pipeline to form a local circulation, and part of the waste solution flows out from the top inlet and outlet of the tangential flow chromatography column and is collected through the liquid outlet main pipeline;

[0021] Step 3: Flushing after loading, including tangential flushing and reverse flushing

[0022] Step 4: Elution, the eluent flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the bottom inlet and outlet, enters the liquid outlet main pipeline and is collected; the eluent flows into the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the top inlet and outlet, enters the liquid outlet main pipeline and is collected.

[0023] Further in some embodiments, two filters are used, and the first inlet and outlet ends of the two filters are respectively connected to the reflux end pipeline and the waste pipeline of the tangential flow chromatography column; the second inlet and outlet ends of the two filters are respectively connected to the liquid inlet main pipeline and the cleaning pipeline. When one of the filters is performing filtering work, the other filter is in backwashing work.

[0024] Further, in some embodiments, the tangential flushing is as follows: the buffer solution enters the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the reflux port, passes through the filter and enters the liquid inlet main pipeline to form a cycle; the reverse flushing is as follows: the buffer solution flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the reflux port, passes through the filter and enters the liquid inlet main pipeline to form a cycle.

[0025] The present invention has the following effects:

[0026] 1. The reflux end of the tangential flow chromatography column used in the present invention is connected with a filter module to prevent clogging of the sieve plate;

[0027] 2. After loading, tangential flushing and reverse flushing can be used. The buffer solution passes through the reflux end of the tangential flow chromatography column and the filter module into the main liquid inlet pipeline, so that the sieve plate can be flushed tangentially and reversely, and is not easy to clog.

[0028] In summary, high linear flow rate and overload loading can be achieved, achieving extremely high dynamic loading capacity. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a structural schematic diagram of the present invention. DETAILED DESCRIPTION

[0030] The present invention will be further described below in conjunction with the accompanying drawings.

[0031] The technical content of the present invention is described below by specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments. Those skilled in the art can make various modifications and changes without departing from the spirit of the present invention.

[0032] Before describing the specific embodiments of the present disclosure in detail, some terms used in the present disclosure are first explained.

[0033] Unless otherwise defined below, the meaning of all technical terms and scientific terms used herein is intended to be the same as those commonly understood by those skilled in the art. Mentioning the technology used in this article is intended to refer to the technology commonly understood in the art, including those changes in technology that are obvious to those skilled in the art or the replacement of equivalent technology. Although it is believed that the following terms are well understood by those skilled in the art, the following definitions are still set forth to better explain the present invention. When the name of a top grade appears in this article, it is intended to refer to its corresponding top grade. All patents, published patent applications and publications cited herein are incorporated herein by reference.

[0034] As used herein, the terms “connected”, “connected”, “coupled” or “coupled” and the like are not limited to direct connections but also include indirect connections.

[0035] As used herein, a "sample" is a biomolecule, including proteins, nucleic acids, lipids, carbohydrates, small nucleotides, amino acids and their derivatives.

[0036] As used herein, the term "online monitoring" or "real-time monitoring" refers to real-time detection of certain parameters or properties of buffers, reaction fluids, and fluids flowing out of flow reactors during use of the chromatography system, such as pH, pressure, flow rate, conductivity, etc. Unlike offline detection or analysis, online monitoring or real-time monitoring can provide real-time feedback of the detection results.

[0037] The position relationship words "up", "down", "left", "right", "front", "back", etc. involved in this article are determined according to the layout direction of the drawings in the specification. They are only used to indicate relative position relationships. When the absolute position of the object being described changes, the relative position relationship may also change accordingly.

[0038] The RTF chromatography column (tangential flow chromatography column) mentioned in this article is a patented chromatography column independently developed by our company, with patent number: 202220697921.8; it has a radial and tangential flow structure, and can be loaded through the tangential flow at the bottom of the column, while the upper part of the column head can be flushed and eluted at the same time, cyclic loading and cleaning mode with equilibrium liquid can be achieved, thereby achieving high linear flow rate and overload loading, and achieving extremely high dynamic loading capacity.

[0039] Reference Figure 1 As shown, a chromatography system based on a tangential flow chromatography column comprises a liquid inlet main pipeline 1, the inlet end of the liquid inlet main pipeline 1 is respectively connected to a variety of solutions, such as a product tank 2, a buffer tank 3, an eluent tank 4 and a cleaning agent tank 5 as shown in the figure, the outlet end pipeline of the product tank 2 is connected to the inlet end of the liquid inlet main pipeline 1 through a valve AV105, the outlet pipeline of the buffer tank 3 is connected to the inlet end of the liquid inlet main pipeline 1 through a valve AV102, the outlet pipeline of the eluent tank 4 is connected to the inlet end of the liquid inlet main pipeline 1 through a valve AV103, and the outlet pipeline of the cleaning agent tank 5 is connected to the inlet end of the liquid inlet main pipeline 1 through a valve AV104. The liquid inlet main pipeline 1 is provided with a liquid inlet pump 6, a first flow indicating transmitter 7, a first pressure sensor 8 and a first conductivity transmitter 9.

[0040] The outlet end of the inlet main pipeline 1 is connected to the first valve array 10, which is a rectangular valve array formed by connecting valves AV401, AV402, AV404 and AV405 end to end. The outlet end of the inlet main pipeline 1 is connected between valves AV401 and AV402. The pipelines of the bottom inlet and outlet of the tangential flow chromatography column 11 are connected between valves AV401 and AV404, the pipelines of the top inlet and outlet of the tangential flow chromatography column 11 are connected between valves AV405 and AV402, the inlet end of the outlet main pipeline 12 is connected between valves AV404 and AV405, and the inlet end of the inlet main pipeline 12 is connected between the valves AV403.

[0041] The main liquid outlet pipeline 12 is provided with a pressure control valve PCV302, a first UV sensor 13, a second flow indicating transmitter 14, a second conductivity transmitter 15 and a first pH sensor 16. The outlet end of the main liquid outlet pipeline 12 is connected to a plurality of collecting ports, such as the product collecting tank 17 and the waste tank 18 as shown in the figure. The inlet end pipeline of the product collecting tank 17 is connected to the outlet end of the main liquid outlet pipeline 12 through the valve AV101, and the inlet pipeline of the waste tank 18 is connected to the outlet end of the main liquid outlet pipeline 12 through the valve AV201.

[0042] The pipeline at the reflux end of the tangential flow chromatography column 11 is respectively provided with a second pressure sensor 19, a valve AV301 and a pressure control valve PCV302.

[0043] The outlet end of the reflux end pipeline of the tangential flow chromatography column 11 is connected to the second valve array 20, and the second valve array 20 is a rectangular valve array formed by connecting valves AV601, AV602, AV603 and AV604 end to end. The outlet end of the reflux end pipeline of the tangential flow chromatography column 11 is connected between valves AV601 and AV603, and a third pressure sensor 21 is arranged between the tangential flow chromatography column 11 and valves AV601 and AV603. The top inlet and outlet ends of the first filter 22 are connected between valves AV601 and AV602. The top inlet and outlet ends of the second filter 23 are connected between valves AV603 and AV604. The waste discharge pipeline is connected between valves AV602 and AV604.

[0044] The inlet and outlet ends at the bottom of the first filter 22 and the second filter 23 are connected to the third valve array 24, which is a rectangular valve array formed by connecting valves AV605, AV606, AV607 and 608 end to end. The inlet and outlet ends at the bottom of the first filter 22 are connected between valves AV605 and AV606; the inlet and outlet ends at the bottom of the second filter 23 are connected between valves AV607 and 608; the inlet end of the WFI solution is connected between valves AV605 and AV607; valves AV606 and 608 are connected to the main liquid inlet pipeline 1 through a pipeline, and the third pressure sensor 25 and valve AV106 are respectively provided on the pipeline, and the connection point between the pipeline and the main liquid inlet pipeline 1 is located on the left side of the liquid inlet pump 6.

[0045] The feed inlet pipeline at the top of the tangential flow chromatography column 11 is connected to the top inlet pipeline of the filling tank 26 and the bottom outlet pipeline of the filling tank 26, respectively. The top inlet pipeline of the filling tank 26 is provided with a valve AV503, and the two ends of the bottom outlet pipeline of the filling tank 26 are provided with valves AV501 and AV502, respectively, and a feed pump 27 between the two valves. The upper half of one side of the filling tank 2 is provided with a liquid outlet pipeline, which is connected to the liquid inlet main pipeline 1 through a valve AV107, and the connection point is located on the left side of the liquid inlet pump 6.

[0046] When used, the steps include:

[0047] Step 1: Balance:

[0048] Open valve AV102 to allow the buffer to flow out of the buffer tank 3 and into the liquid inlet main pipeline 1. The first flow indicating transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor data in real time. Open valve AV402 to allow the buffer to enter from the top inlet and outlet of the tangential flow chromatography column 11 and flow out from the bottom inlet and outlet of the tangential flow chromatography column 11. Open valve AV404 to allow the buffer to flow into the liquid outlet main pipeline 12. The first UV sensor 13, the second flow indicating transmitter 14, the second conductivity transmitter 15, and the first pH sensor 16 monitor data in real time. Finally, the buffer enters the waste tank 18.

[0049] Step 2: Sample loading

[0050] Open valve AV105, the sample solution to be processed flows out of the product tank 2 into the liquid inlet main pipeline 1, the first flow indicating transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time, open valve AV401 to flow in from the bottom inlet and outlet of the tangential flow chromatography column 11, and part of the solution flows out from the reflux end of the tangential flow chromatography column 11. At this time, open valve AV301, valve AV601 on the second valve array 20, valve AV606 on the third valve array 24, and valve AV106. At this time, the solution flows in through the top inlet and outlet of the first filter 22, and flows out from the bottom inlet and outlet of the first filter 22 into the liquid inlet main pipeline 1, forming a cycle. At the same time, the second filter 23 can be flushed. At this time, the valve AV604 in the second valve array 20 and the valve 607 in the third valve array 24 are opened. At this time, the WFI solution enters the inlet and outlet at the bottom of the second filter 23 through the WFI solution inlet end, and then flows into the waste discharge pipeline from the inlet and outlet ends at the top of the second filter 23; similarly, when the second filter 23 is performing filtering work, the valve AV603 in the second valve array 20 and the valve AV608 in the third valve array 24 are opened. At this time, the solution flows in through the top inlet and outlet ends of the second filter 23, and flows out from the bottom inlet and outlet ends of the second filter 23 into the liquid inlet main pipeline 1, forming a cycle; the first filter 22 is backwashed, and the valve AV602 in the second valve array 20 and the valve 605 in the third valve array 24 are opened. At this time, the WFI solution enters the inlet and outlet ends at the bottom of the first filter 22 through the WFI solution inlet end, and then flows into the waste discharge pipeline from the inlet and outlet ends at the top of the first filter 22; in this way, the two filters can be used alternately.

[0051] The solution entering the tangential flow chromatography column 11 passes through the inlet and outlet at the top of the tangential flow chromatography column 11, opens the valve AV405, enters the liquid outlet main pipeline 12, and the first UV sensor 13, the second flow indicating transmitter 14, the second conductivity transmitter 15 and the first pH sensor 16 monitor the data in real time, and finally enters the waste tank 18.

[0052] Step 3: Post-sampling flushing, including tangential flushing and back flushing

[0053] 1. Tangential flushing: Open valve AV102 and the buffer solution flows out of the buffer solution tank 3 and enters the liquid inlet main pipeline 1. The first flow indicator transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time. Open valve AV401 and flow into the bottom inlet and outlet of the tangential flow chromatography column 11. The solution flows out from the reflux end of the tangential flow chromatography column 11, passes through the first filter 22 or the second filter 23, and flows into the liquid inlet main pipeline 1 to form a cycle.

[0054] 2. Back flushing: Open valve AV102 and the buffer solution flows out of the buffer solution tank 3 and enters the liquid inlet main pipeline 1. The first flow indicator transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time. Open valve AV402 and the buffer solution flows into the top inlet and outlet of the tangential flow chromatography column 11, flows out from the reflux end of the tangential flow chromatography column 11, passes through the first filter 22 or the second filter 23, and flows into the liquid inlet main pipeline 1 to form a cycle.

[0055] Step 3: Elution is divided into top-in and bottom-out and top-in and bottom-out

[0056] 1. In from the top and out from the bottom: open valve AV103, the eluent flows out from the eluent tank 4 and enters the liquid inlet main pipeline 1, the first flow indicating transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time, open valve AV402 to allow the eluent to enter from the top inlet and outlet of the tangential flow chromatography column 11, and the solution flows out from the bottom inlet and outlet of the tangential flow chromatography column 11, open valve AV404 and enter the liquid outlet main pipeline 12, the first UV sensor 13, the second flow indicating transmitter 14, the second conductivity transmitter 15 and the first pH sensor 16 monitor the data in real time, and finally enter the product collection tank 17.

[0057] 2. In from the top and out from the bottom: open valve AV103, the eluent flows out from the eluent tank 4 and enters the liquid inlet main pipeline 1, the first flow indicating transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time, open valve AV401, and allow the eluent to enter from the bottom inlet and outlet of the tangential flow chromatography column 11, and the solution flows out from the top inlet and outlet of the tangential flow chromatography column 11, open valve AV405 and enter the liquid outlet main pipeline 12, the first UV sensor 13, the second flow indicating transmitter 14, the second conductivity transmitter 15 and the first pH sensor 16 monitor the data in real time, and finally enter the product collection tank 17.

[0058] Step 4: Regeneration, open valve AV104, the cleaning agent flows out from the cleaning agent tank 5 and enters the liquid inlet main pipeline 1, the first flow indicating transmitter 7, the first pressure sensor 8, and the first conductivity transmitter 9 monitor the data in real time, open valve AV401, so that the cleaning agent enters from the bottom inlet and outlet of the tangential flow chromatography column 11, and the solution flows out from the top inlet and outlet of the tangential flow chromatography column 11, open valve AV405 and enter the liquid outlet main pipeline 12, the first UV sensor 13, the second flow indicating transmitter 14, the second conductivity transmitter 15 and the first pH sensor 16 monitor the data in real time, and finally enter the waste tank 18.

[0059] Step 5: Loading the filler. At this time, open valve AV501, valve AV502 and feed pump 27 on the bottom outlet pipeline of the filler tank 26, and the filler enters the tangential flow chromatography column 11 from the feed port at the top of the tangential flow chromatography column 11.

[0060] Step six: unloading the filler. At this time, valve AV107 is closed, and the upper solution in the filler tank 26 enters the liquid inlet main pipeline 1 through the liquid inlet pump 6, and then passes through valve AV401, enters the column from the bottom inlet and outlet of the tangential flow chromatography column 11, and pushes the filler out of the column, and then flows into the top inlet pipeline of the filler tank 26 through the feed inlet pipeline at the top of the tangential flow chromatography column 11, and the filler enters the filler tank 26 to complete the recovery.

Claims

1. A chromatography system based on a tangential flow chromatography column, characterized in that: It comprises a liquid inlet main pipeline and a liquid outlet main pipeline; the liquid inlet main pipeline is provided with a liquid inlet pump; the outlet end of the liquid inlet main pipeline is connected to the inlet and outlet pipelines at the bottom and top of the tangential flow chromatography column through valves; the inlet end of the liquid outlet main pipeline is connected to the inlet and outlet pipelines at the bottom and top of the tangential flow chromatography column through valves; the reflux end pipeline of the tangential flow chromatography column is connected to the liquid inlet main pipeline through a connecting pipeline, and a filtering module is provided on the connecting pipeline.

2. A chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: A first valve array is provided at the outlet end of the liquid inlet main pipeline, and the first valve array includes a rectangular valve array composed of four valves connected at the ends. The outlet end of the liquid inlet main pipeline and the inlet end of the liquid outlet main pipeline are respectively connected to a group of opposite sides of the rectangular valve array; the other group of opposite sides of the rectangular valve array are respectively connected to the inlet and outlet pipelines of the corresponding tangential flow chromatography column bottom and top.

3. A chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The filtration module is a filter, the pipelines at the first inlet and outlet ends of the filter are connected to the reflux end pipeline of the tangential flow chromatography column, and the pipelines at the second inlet and outlet ends of the filter are connected to the liquid inlet main pipeline.

4. A chromatography system based on a tangential flow chromatography column according to claim 3, characterized in that: The pipelines at the first inlet and outlet ends of the filter are connected to the waste pipeline, and the pipelines at the second inlet and outlet ends of the filter are connected to the cleaning pipeline.

5. A chromatography system based on a tangential flow chromatography column according to claim 4, characterized in that: The filter adopts two, the first inlet and outlet ends of the two filters are respectively connected to the reflux end pipeline and the waste pipeline of the tangential flow chromatography column; the second inlet and outlet ends of the two filters are respectively connected to the liquid inlet main pipeline and the cleaning pipeline.

6. A chromatography system based on a tangential flow chromatography column according to claim 5, characterized in that: The first inlet and outlet ends and the second inlet and outlet ends of the two filters are respectively provided with a second valve array and a third valve array. The second valve array is a rectangular valve array composed of four valves connected at the ends. The reflux end pipeline and the waste discharge pipeline of the tangential flow chromatography column are respectively connected to a group of opposite sides of the rectangular valve array, and the other group of opposite sides of the rectangular valve array are respectively connected to the first inlet and outlet ends of the corresponding two filters; the third valve array is a rectangular valve array composed of four valves connected at the ends. One group of opposite sides of the rectangular valve array is respectively connected to the second inlet and outlet ends of the corresponding two filters, and the other group of opposite sides of the rectangular valve array are respectively connected to the corresponding cleaning pipeline and the liquid inlet main pipeline.

7. A chromatography system based on a tangential flow chromatography column according to claim 6, characterized in that: Pressure sensors are respectively arranged between the reflux end pipeline of the tangential flow chromatography column and the second valve array, and on the connecting pipeline between the third valve array and the liquid inlet main pipeline.

8. The chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The top of the tangential flow chromatography column is provided with a feed end, and the pipelines of the feed end are respectively connected to the top and bottom inlet and outlet pipelines of the filling tank, and the inlet and outlet pipelines are respectively provided with valves; the bottom inlet and outlet pipelines are provided with a feed pump.

9. A chromatography system based on a tangential flow chromatography column according to claim 8, characterized in that: A liquid outlet is provided on one side of the stuffing tank, and the liquid outlet is connected to a liquid inlet main pipeline through a pipeline, and a valve is provided on the pipeline.

10. A chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The liquid inlet main pipeline is provided with a flow indicating transmitter and / or a pressure sensor and / or a conductivity transmitter.

11. A chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The liquid outlet main pipeline is provided with a pH sensor and / or a conductivity transmitter and / or a flow indicating transmitter and / or a UV sensor.

12. A chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: A pressure sensor is arranged on the reflux end pipeline of the tangential flow chromatography column.

13. A method for using a chromatography system based on a tangential flow chromatography column, characterized in that: Step 1: Balance, the buffer solution flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the bottom inlet and outlet, enters the liquid outlet main pipeline and is collected; Step 2: Loading the sample, the sample solution to be processed flows into the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, part of the sample solution enters the filtration module from the reflux port, and then enters the liquid inlet main pipeline to form a local circulation, and part of the waste solution flows out from the top inlet and outlet of the tangential flow chromatography column and is collected through the liquid outlet main pipeline; Step 3: Flushing after loading, including tangential flushing and reverse flushing Step 4: Elution, the eluent flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the bottom inlet and outlet, enters the liquid outlet main pipeline and is collected; the eluent flows into the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the top inlet and outlet, enters the liquid outlet main pipeline and is collected.

14. The method for using a chromatography system based on a tangential flow chromatography column according to claim 13, characterized in that: The filters are two, the first inlet and outlet ends of the two filters are respectively connected to the reflux end pipeline and the waste discharge pipeline of the tangential flow chromatography column; the second inlet and outlet ends of the two filters are respectively connected to the liquid inlet main pipeline and the cleaning pipeline. When one of the filters is filtering, the other filter is in backwashing operation.

15. A method for using a chromatography system based on a tangential flow chromatography column according to claim 13, wherein the tangential flushing is: the buffer solution enters the bottom inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the reflux port, passes through the filter and enters the liquid inlet main pipeline to form a cycle; the reverse flushing is: the buffer solution flows into the top inlet and outlet of the tangential flow chromatography column through the liquid inlet main pipeline, flows out from the reflux port, passes through the filter and enters the liquid inlet main pipeline to form a cycle.

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