Continuous chromatographic system based on tangential flow chromatographic column

Abstract

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

Description

TECHNICAL FIELD

[0001] The utility model relates to biological medicine downstream separation and purification, especially based on tangential flow chromatography column's continuous chromatography system. BACKGROUND

[0002] The chromatography column of the chromatography system downstream of biological medicine is re-used from the structure that the sample to be treated enters from one end of the chromatography column and flows out from the other end to be collected, so that it cannot handle particle precipitates and cannot backflush the distribution screen plate, and cannot realize the ability of simultaneous high-efficiency chromatography and particle treatment. In the prior art, for example, patent number 202120067489.X, the utility model name is a hydraulic drive type dynamic chromatography device. When the separation liquid needs to be added, the liquid is injected from the liquid inlet and flows out from the liquid outlet to be collected. Or adopt for example patent number 201820028656.8, the utility model name is a dynamic beam type full-automatic chromatography column. The structure adopted at the bottom is a sieve plate, a flow distribution plate and a base. The center position of the sieve plate is provided with a lower spray head. When the separation liquid needs to be added, the liquid enters from the chromatography port of the upper spray head and flows out from the chromatography port of the lower spray head to be collected, or enters from the chromatography port of the lower spray head and flows out from the chromatography port of the upper spray head to be collected. The disadvantages are: 1. Cannot realize column head upper washing or elution; 2. Does not support reverse flushing of the filter plate (sieve plate), which is easy to cause blockage; 3. Cannot realize high linear flow rate and overload sample loading at the same time, and the dynamic load is low.

[0003] The applicant proposes a tangential flow chromatography column, patent number 202220697921.8, patent name a radial, tangential flow bottom for chromatography column and its combined chromatography system. Adopting tangential flow structure, realizing chromatography sample loading while tangential flow filtration, realizing high linear flow rate and overload sample loading, realizing extremely high dynamic load. Therefore, in order to meet the production demand, it is necessary to use the continuous flow chromatography system of tangential flow chromatography column. UTILITY MODEL CONTENT

[0004] In order to achieve the above purpose, the utility model provides a continuous chromatography system based on tangential flow chromatography column, which can realize that part of the chromatography column is in loading mode and part is 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 purpose, the utility model adopts the technical scheme that:

[0006] The utility model discloses a first aspect provides a kind of continuous chromatography system based on tangential flow chromatography column, including sample introduction pipeline, elution pipeline and multiple tangential flow chromatography column;The sample introduction pipeline is provided with sample introduction pump, and the outlet end of the sample introduction pipe is respectively connected with the inlet and outlet end pipeline of the bottom of each tangential flow chromatography column by valve respectively;The elution pipeline is provided with elution pump, and the outlet end of the elution pipeline is respectively connected with the inlet and outlet end pipeline of the bottom of each tangential flow chromatography column by valve;The inlet and outlet end pipeline of the top of each tangential flow chromatography column is respectively connected with first collection pipeline and second collection pipeline by valve;The return port pipeline of the tangential flow chromatography column is connected with the sample introduction pipeline by filter module or liquid storage tank.

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

[0008] In some embodiments, the outlet end pipeline of the sample introduction pipeline and elution pipeline is respectively connected with first valve array;The first valve array is rectangular valve array formed by four ends connected;One pair of sides of rectangular valve array is respectively connected with the outlet end of sample introduction pipeline and elution pipeline;Another pair of sides of rectangular valve array is respectively connected with the bottom inlet and outlet end of two chromatography columns.

[0009] In some embodiments, the top inlet and outlet end pipeline of the two is respectively connected with second valve array, and the first valve array is rectangular valve array formed by four ends connected;One pair of sides of rectangular valve array is respectively connected with the top inlet and outlet end pipeline of two, and another pair of sides of rectangular valve array is respectively connected with the inlet end of first collection pipeline and second collection pipeline.

[0010] In some embodiments, the filter module adopts multiple filters, and the inlet and outlet of each filter are respectively connected with the return port end of two tangential flow chromatography columns by valve, and the other end of filter is connected with sample introduction pipeline.

[0011] In some embodiments, the filter adopts two.

[0012] In some embodiments, the return end pipeline of the two tangential flow chromatography columns is connected with two filters by third valve array, and the third valve array is rectangular valve array formed by four ends connected;One pair of sides of rectangular valve array is respectively connected with the inlet and outlet pipeline of one end of two filters;Another pair of sides of rectangular valve array is respectively connected with return end pipeline and waste pipe line;The other end of two filters is connected with sample introduction pipeline by fourth valve array, and the fourth valve array is rectangular valve array formed by four ends connected;One pair of sides of rectangular valve array is respectively connected with the inlet and outlet pipeline of the other end of two filters, and another pair of sides of rectangular valve array is respectively connected with liquid inlet pipeline and sample introduction pipeline.

[0013] In some embodiments, a recirculation pump is provided on the recirculation port line of the tangential flow chromatography column.

[0014] In some embodiments, the outlet line of the filter module or the reservoir is connected to the sample inlet line through a mixer.

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

[0016] The beneficial effects of the present application are as follows:

[0017] 1. Two tangential flow chromatography columns are used in the present application, which are operated alternately: one is in loading mode, and the other is in elution mode. By doing so, the feed can be continuously loaded into the RTF column, and there is almost no residual in the flow path and the circulation loop, and the circulation loop does not have to be stopped and restarted multiple times.

[0018] 2. The recirculation end of the tangential flow is connected to the circulation loop, which drives the circulation while loading the tangential flow chromatography column.

[0019] 3. The filter module is used to remove particulate matter or contaminants from the feed and maintain a constant particulate matter concentration (lower than that of the feed). BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The structure of the present application is shown in the schematic diagram. DETAILED DESCRIPTION

[0021] The present application will be further described below with reference to the accompanying drawings.

[0022] The technical content of the present application is described below through specific embodiments, and those skilled in the art can easily understand other advantages and effects of the present application from the disclosed content of the present application. The present application 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 application.

[0023] Before describing the specific embodiments of the present disclosure in detail, first, some terms used in the present disclosure are explained.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Reference herein to techniques used in the present application are intended to refer to those techniques generally understood by those in the art, including variations or substitutions of techniques that would be apparent to one of ordinary skill in the art. Although the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate a better understanding of the present application. When an item is referred to herein, it is intended to refer to its counterpart as well. All patents, published patent applications, and publications recited herein are incorporated by reference herein.

[0025] The terms "connected," "coupled," or "coupling," and like terms as used herein, are not intended to be limited to direct connections only.

[0026] A "sample" as described herein is a biomolecule, including proteins, nucleic acids, lipids, carbohydrates, small nucleotides, amino acids, and derivatives thereof.

[0027] The terms "on-line monitoring" or "real-time monitoring" as used herein refer 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.

[0028] 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, and are only used to represent the relative positional relationship. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

[0029] The RTF chromatography column (tangential flow chromatography column) as referred to herein is a chromatography column independently developed by the company and has a patent, the patent number is: 202220697921.8; has a radial and tangential flow structure, can be loaded through the tangential flow at the bottom of the column, and at the same time the upper part of the column head can be washed and eluted, and the cycle loading and washing mode with the balance liquid is used, so as to realize high linear flow rate and overload loading, and realize extremely high dynamic capacity.

[0030] Referring to Figure 1As shown, a continuous chromatography system based on tangential flow chromatography column, including sample loading pipeline 1, elution pipeline 2; the inlet end of sample loading pipeline 1 is connected with first tank 3 through valve AV1, and the first tank 3 can be used for loading sample to be treated. The inlet end of sample loading pipeline 1 is connected with second tank 4 through valve AV4. Sample loading pump 5 and first flow indicating transmitter 6 are arranged on sample loading pipeline 1. The outlet end of sample loading pipeline 1 is connected with first valve array 7, and the first valve array 7 is a rectangular valve array composed of valve AV10, valve AV11, valve AV12 and valve AV13 connected in a head-to-tail manner, and the sample loading pipeline 1 is connected between valve AV10 and valve AV11. The inlet end of elution pipeline 2 is connected with second tank 5 through valve AV5, third tank 8 through valve AV2 and fourth tank 9 through valve AV3. Elution pump 10, second flow indicating transmitter 11 and bubble trap 33 are arranged on elution pipeline 2. The outlet end of elution pipeline 2 is connected between valve AV12 and valve AV13 through a pipeline, and valve AV14 is arranged on the pipeline. The bottom inlet and outlet ends of first tangential flow chromatography column 12 are connected between valve AV10 and valve AV12 through a pipeline; and the bottom outlet end of second tangential flow chromatography column 13 is connected between valve AV11 and valve AV13 through a pipeline.

[0031] The top inlet and outlet ends of first tangential flow chromatography column 12 and second tangential flow chromatography column 13 are connected with second valve array 14, and the second valve array 14 is a rectangular valve array composed of valve AV17, valve AV18, valve AV19 and valve AV20 connected in a head-to-tail manner. The pipeline connected with the top inlet and outlet ends of first tangential flow chromatography column 12 is connected between valve AV19 and valve AV20, and the pipeline connected with the top inlet and outlet ends of second tangential flow chromatography column 13 is connected between valve AV17 and valve AV18. First collection pipeline 15 is connected between valve AV17 and valve AV19; and second collection pipeline 16 is connected between valve AV18 and valve AV20. First UV sensor 17, first conductivity transmitter 18 and first pH sensor 19 for monitoring real-time data are arranged on first collection pipeline 15. Second UV sensor 20, second conductivity transmitter 21 and second pH sensor 22 for monitoring real-time data are arranged on second collection pipeline 16.

[0032] The return port pipeline of the first tangential flow chromatography column 12 is connected to the inlet end of the connecting pipeline 23 through the valve AV6, and the return port pipeline of the second tangential flow chromatography column 13 is connected to the connecting pipeline 23 through the valve AV7. The connecting pipeline 23 is provided with a circulating pump 24 and a third flow indicating transmitter 25. The outlet end of the connecting pipeline 23 is connected to a third valve array 26, which is a rectangular valve array composed of a pressure control valve PCV1, a pressure control valve PCV2, a valve AV26 and a valve AV27 connected end to end. The connecting pipeline 23 is connected between the pressure control valve PCV1 and the pressure control valve PCV2. The first inlet and outlet end pipeline of the first filter 27 is connected between the pressure control valve PCV1 and the valve AV26. The first inlet and outlet end pipeline of the second filter 28 is connected between the pressure control valve PCV2 and the valve AV27. The waste pipeline 29 is connected between the valve AV26 and the valve AV27. The second inlet and outlet end of the first filter 27 and the second filter 28 is connected to a fourth valve array 30, which is a rectangular valve array composed of a valve AV22, a valve AV23, a valve AV24 and a valve AV25 connected end to end. The second inlet and outlet end pipeline of the first filter 27 is connected between the valve AV22 and the valve AV23. The second inlet and outlet end pipeline of the second filter 28 is connected between the valve AV24 and the valve AV25. The liquid inlet pipeline 31 is connected between the valve AV22 and the valve AV24. The sample loading pipeline 1 is connected between the valve AV23 and the valve AV25. The valve AV21 is arranged on the left side of the connection point of the pipeline and the sample loading pipeline 1.

[0033] The outlet end of the elution pipeline 2 is connected to the top inlet and outlet end pipelines of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 through pipelines, respectively, and the pipelines are respectively provided with the valve AV16 and the valve AV15.

[0034] The pipelines of the bottom return ports of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13 are respectively connected to the solution collection pipeline through the valve AV18 and the valve AV19.

[0035] The sample loading pipeline 1 is provided with a mixer 32 between the fourth valve array 30.

[0036] 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 simultaneously eluted or one is loaded and the other is eluted.

[0037] 1、When loading simultaneously: the sample to be treated flows out from the first tank 3, enters the loading pipeline 1 under the action of the loading pump 5, the valve AV10 and the valve AV11 are opened, the sample to be treated enters the bottom inlet and outlet 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 backflow pipelines at the bottom 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.

[0038] The filter module adopts the first filter 27 and the 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.

[0039] At this time, the pressure control valve PCV1 and the valve AV23 are opened, the solution flows in through the first inlet and outlet ends of the first filter 27 and flows out from the second inlet and outlet ends of the first filter 27, and finally flows into the 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 inlet and outlet pipelines at the top of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, at this time, the valve AV19 and the valve AV17 are opened, and the waste solution enters the first collection pipeline 15 for collection.

[0040] The second filter 28 performs backwashing: the valve AV24 and the valve AV27 are opened, the WFI solution enters the second inlet and outlet ends of the second filter 28 through the valve AV24, after backwashing, the waste solution enters the waste pipeline 29 after passing through the valve AV27.

[0041] 2、When eluting simultaneously: the eluent enters the elution pipeline 2 under the action of the elution pump 10, the valve AV12 and the valve AV13 are opened, 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, passes through the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, and the solution flows out from the inlet and outlet pipelines at the top of the first tangential flow chromatography column 12 and the second tangential flow chromatography column 13, at this time, the valve AV20 and the valve AV18 are opened, and the solution enters the second collection pipeline 16 for collection.

[0042] 3. One in loading and one in elution: (1) The first tangential flow chromatography column 12 is loaded and the second tangential flow chromatography column 13 is eluted: The sample to be treated 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 treated enters the bottom inlet and outlet end pipeline of the first tangential flow chromatography column 12. Part of the solution enters the connecting pipeline 23 through the backflow end pipeline at the bottom of the first tangential flow chromatography column 12, enters the filtering module through the connecting pipeline 23, and the filtering module uses the first filter 27 and the second filter 28. The two filters can perform filtering work at the same time or one performs filtering work and the other performs backwashing; or they can filter at the same time or backwash at the same time. For example, the first filter 27 is backwashed and the second filter 28 is filtered. At this time, the pressure control valve PCV2 and the valve AV25 are opened, the solution flows in through the first inlet and outlet end of the second filter 28, flows out from the second inlet and outlet end of the second filter 28, and finally flows into the loading pipeline 1 to form a circulation. The sample to be treated passes through the first tangential flow chromatography column 12, and the waste 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 AV19 is opened, and the waste solution enters the first collection pipeline 15 for collection. The first filter 27 is backwashed: the valve AV22 and the valve AV26 are opened, and the WFI solution enters the second inlet and outlet end of the first filter 27 from the valve AV22 for backwashing. After backwashing, the waste solution enters the waste pipeline 29 through the valve AV26.

[0043] At the same time, the eluent enters the elution pipeline 2 under the action of the elution pump 10. The valve AV13 is opened, the eluent enters the bottom inlet and outlet end pipeline of the second tangential flow chromatography column 13, passes through the column of the second tangential flow chromatography column 13, and the 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 AV18 is opened, and the solution enters the second collection pipeline 16 for collection.

[0044] (2) The first tangential flow chromatography column 12 is eluted and the second tangential flow chromatography column 13 is loaded: The sample to be treated flows out from the first tank 3 and enters the loading pipeline 1 under the action of the loading pump 5. The valve AV11 is opened, and the sample to be treated enters the bottom inlet and outlet end pipeline of the second tangential flow chromatography column 13. Part of the solution enters the connecting pipeline 23 through the backflow end pipeline at the bottom of the second tangential flow chromatography column 13, and enters the filtering module through the connecting pipeline 23. The filtering module uses the first filter 27 and the second filter 28. The two filters can perform filtering work at the same time or one performs filtering work and the other performs backwashing; or they can filter at the same time or backwash at the same time. For example, the first filter 27 is filtered and the second filter 28 is backwashed.

[0045] 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, flows out of the second inlet and outlet end 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 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.

[0046] The second filter 28 is back-flushed: 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, after back-flushing, the waste solution enters the waste pipeline 29 after passing through the valve AV27.

[0047] 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 inlet and outlet end pipeline at the bottom of the first tangential flow chromatography column 12, 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; 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.

2. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The tangential flow chromatography columns are two, one in 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 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 are connected 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 are connected to the inlet and outlet end pipelines at the bottom of the two chromatography columns respectively.

4. 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 chromatography columns are connected to a second 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 are connected to the inlet and outlet end pipelines at the top of the two chromatography columns respectively; and the other pair of opposite sides of the rectangular valve array are connected to the inlet end of the first collection pipeline and the second collection pipeline respectively.

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

6. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The filters are two.

7. A continuous chromatography system based on a tangential flow chromatography column according to claim 6, 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; 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 are connected to the inlet and outlet end pipelines of one end of the two filters respectively; the other pair of opposite sides of the rectangular valve array are connected to the backflow end pipelines and a waste 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 are connected 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 are connected to a liquid inlet pipeline and the sample loading pipeline respectively.

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

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

10. A continuous chromatography system based on a tangential flow chromatography column according to claim 1, characterized in that: The outlet end of the elution pipeline is connected to the liquid inlet end pipelines at the top of the plurality of tangential flow chromatography columns through valves respectively.

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