Protein chromatography, purification and detection integrated device

Abstract

The utility model provides a protein chromatography, purification and detection integrated device which comprises a first pump body, a second pump body, a chromatographic column, an ICE analysis module, a plurality of valves and a plurality of sensors, and the output end of the first pump body converges to a main pipeline through a first branch pipeline; the first output end of the second pump body converges to the main pipeline through a second branch pipeline; the chromatographic column is connected to the main pipeline through a column position valve, the input end of the chromatographic column is connected with a first bubble trap, and the output end of the chromatographic column is connected with the ICE analysis module through a sanitary two-position three-way reversing valve. According to the scheme, full-automatic protein chromatography is carried out through the chromatography flow path combination, and protein chromatography purification is carried out on line in real time; the ICE analysis module is switched through the three-way reversing valve, qualitative analysis can be synchronously carried out, the analysis speed is increased, and multiple purposes are achieved through one machine; moreover, the ICE analysis module is adopted to detect the protein on line, so that the detection time can be greatly shortened, the product activity is reserved to the maximum extent, and a more reliable quality guarantee is provided for biopharmaceutical production.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of biopharmaceutical purification separation technology, especially to a protein chromatography purification detection integrated device. BACKGROUND

[0002] Traditional protein chromatography purification analysis is often operated after a series of operations such as small test or production, in order to understand the purity and properties of the product, the protein sample after chromatography purification needs to be sent to QC inspection and detection. There will be waiting time in the detection process, and denaturation and activity attenuation may occur after production of some specific proteins, the faster the freeze-drying packaging, the better the product activity and effect can be guaranteed, so the test result needs to be determined as soon as possible to decide whether to freeze-dry and package, reprocess or scrap.

[0003] However, the existing protein chromatography purification and detection analysis after protein chromatography purification often need two sets of equipment to operate respectively, and part of the time is consumed in transferring the protein sample; and the detection analysis method applied to the protein chromatography purification usually adopts gel plate isoelectric focusing detection, the traditional gel plate isoelectric focusing detection takes a long time, and the time consumed by multiple groups of small tests may also cause loss of the activity of specific proteins. Therefore, it is necessary to provide a protein chromatography purification detection integrated device to overcome the above problems. SUMMARY

[0004] The utility model aims at solving the above problems, and provides a protein chromatography purification detection integrated device to solve the problem of long process consumption time between protein chromatography purification and detection analysis after purification, which easily leads to protein denaturation or inactivation.

[0005] The utility model discloses a technical solution is: a protein chromatography purification detection integration device, including first pump body, second pump body, chromatography column and ICE analysis module, first pump body sets up on first branch pipeline, second pump body sets up on second branch pipeline, chromatography column with ICE analysis module all set up on main pipeline, its characteristics are: the output of first pump body is converged to main pipeline through first branch pipeline, and the input of first pump body is connected air flow meter, first liquid inlet valve group in proper order, and the output of first pump body is connected first pneumatic diaphragm valve. The first output of second pump body is converged to main pipeline through second branch pipeline, and the input of second pump body is connected second liquid inlet valve group, and the first output of second pump body is connected mass flowmeter, second pneumatic diaphragm valve in proper order, and the second output of second pump body is connected first normal pressure discharge valve. Chromatography column connects several column position valves, and the input of chromatography column is connected mixer, pressure sensor, first bubble trap and first sensor group in proper order, and the output of chromatography column is controlled by seventh pneumatic diaphragm to liquid, and is connected liquid outlet valve group and ICE analysis module through sanitary grade two position three way reversing valve respectively.

[0006] Further, the protein chromatography purification detection integration device, wherein: the first bubble trap is connected to the main pipeline through the fourth pneumatic diaphragm valve, and the first bubble trap is provided with a liquid inlet port, a liquid outlet port and an exhaust port, a third pneumatic diaphragm valve is arranged on the liquid inlet port, a fifth pneumatic diaphragm valve is arranged on the liquid outlet port, and a sixth pneumatic diaphragm valve is arranged on the exhaust port.

[0007] Further, the protein chromatography purification detection integration device, wherein: the first bubble trap is connected to the main pipeline through the fourth pneumatic diaphragm valve, and the first bubble trap is provided with a liquid inlet port, a liquid outlet port and an exhaust port, a third pneumatic diaphragm valve is arranged on the liquid inlet port, a fifth pneumatic diaphragm valve is arranged on the liquid outlet port, and a sixth pneumatic diaphragm valve is arranged on the exhaust port.

[0008] Further, the protein chromatography purification detection integration device, wherein: the first bubble trap is connected to the main pipeline through the fourth pneumatic diaphragm valve, and the first bubble trap is provided with a liquid inlet port, a liquid outlet port and an exhaust port, a third pneumatic diaphragm valve is arranged on the liquid inlet port, a fifth pneumatic diaphragm valve is arranged on the liquid outlet port, and a sixth pneumatic diaphragm valve is arranged on the exhaust port.

[0009] Further, the protein chromatography purification detection integration device, wherein: the first bubble trap is connected to the main pipeline through the fourth pneumatic diaphragm valve, and the first bubble trap is provided with a liquid inlet port, a liquid outlet port and an exhaust port, a third pneumatic diaphragm valve is arranged on the liquid inlet port, a fifth pneumatic diaphragm valve is arranged on the liquid outlet port, and a sixth pneumatic diaphragm valve is arranged on the exhaust port.

[0010] Further, the protein chromatography purification detection integration device, wherein: the first bubble trap is connected to the main pipeline through the fourth pneumatic diaphragm valve, and the first bubble trap is provided with a liquid inlet port, a liquid outlet port and an exhaust port, a third pneumatic diaphragm valve is arranged on the liquid inlet port, a fifth pneumatic diaphragm valve is arranged on the liquid outlet port, and a sixth pneumatic diaphragm valve is arranged on the exhaust port.

[0011] Further, the protein chromatography purification and detection integrated device, wherein: the second bubble trap is internally provided with a defoaming zone and a gas-liquid mixing zone, the defoaming zone is an adjacent zone of the liquid inlet port, and the gas-liquid mixing zone is an adjacent zone of the gas outlet port.

[0012] Further, the protein chromatography purification and detection integrated device, wherein: the cross-sectional area of the defoaming zone is smaller than that of the gas-liquid mixing zone.

[0013] Compared with the prior art, the technical scheme of the present application can realize full-automatic protein chromatography through chromatography flow path combination, complete online real-time protein chromatography purification operation, switch to the ICE analysis module through the sanitary two-position three-way directional valve, and simultaneously perform online real-time protein qualitative analysis, thereby improving the analysis speed, meeting various application scenarios from small-scale development to pilot-scale amplification, realizing one machine with multiple functions, and greatly shortening the detection time through online protein detection by the ICE analysis module, synchronously analyzing the product property results in the protein chromatography purification collection time period, maximizing the product activity, and providing more reliable quality guarantee for biopharmaceutical production.

[0014] In addition, two groups of bubble trap structures are arranged, one group of bubble traps is used for defoaming operation before chromatography operation, and the other group of bubble traps is used for secondary defoaming treatment. In this way, on the one hand, the bubbles generated in a section of main pipeline between the first bubble trap and the chromatography column can be effectively removed, and on the other hand, secondary bubble removal is realized, the defoaming efficiency is improved, and the protein chromatography purification quality is improved. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 It is a structural schematic diagram of the whole utility model;

[0016] Figure 2 It is a structural schematic diagram of the chromatography column part of the utility model.

[0017] The meanings of the labels in the attached figures are as follows: 1-Main pipeline, 2-First branch pipeline, 3-Second branch pipeline, 4-First pump body, 5-Second pump body, 6-First bubble trap, 7-Chromatography column, 71-Column body, 72-Second bubble trap, 721-Defoaming zone, 722-Gas-liquid mixing zone, 73-Second liquid level sensor, 74-Inlet port, 75-Outlet port, 76-Exhaust port, 8-ICE analysis module, 9-First inlet valve group, 10-First pneumatic diaphragm valve, 11-Second inlet valve group, 12-Second pneumatic diaphragm valve, 13-First atmospheric pressure exhaust. 14-Third pneumatic diaphragm valve, 15-Fourth pneumatic diaphragm valve, 16-Fifth pneumatic diaphragm valve, 17-Sixth pneumatic diaphragm valve, 18-Second atmospheric pressure discharge valve, 19-First column position valve, 20-Second column position valve, 21-Third column position valve, 22-Fourth column position valve, 23-Seventh pneumatic diaphragm, 24-Sanitary two-position three-way directional valve, 25-Discharge valve assembly, 26-Air flow meter, 27-Mass flow meter, 28-Mixer, 29-Pressure sensor, 30-First liquid level sensor, 31-First sensor assembly, 32-Second sensor assembly. Detailed Implementation

[0018] The technical solution of this utility model will be further described below with reference to the accompanying drawings to make it easier to understand and master. The pump body, valves, and sensors involved are all commonly used components that are generally recognized by those skilled in the art, and there are no special requirements for them in this application.

[0019] like Figure 1 As shown, this utility model provides an integrated protein chromatography purification and detection device, including a first pump body 4, a second pump body 5, a chromatography column 7, and an ICE analysis module 8. The first pump body 4 is disposed on a first branch line 2, the second pump body 5 is disposed on a second branch line 3, and the chromatography column 7 and the ICE analysis module 8 are both disposed on the main line 1.

[0020] According to the technical scheme of the utility model, the output end of the first pump body 4 is converged to the main pipeline 1 through the first branch pipeline 2, the input end of the first pump body 4 is connected with an air flow meter 26 and a first liquid inlet valve group 9 in sequence, and the output end of the first pump body 4 is connected with a first pneumatic diaphragm valve 10. The first output end of the second pump body 5 is converged to the main pipeline 1 through the second branch pipeline 3, the input end of the second pump body 5 is connected with a second liquid inlet valve group 11, the first output end of the second pump body 5 is connected with a mass flow meter 27 and a second pneumatic diaphragm valve 12 in sequence, and the second output end of the second pump body 5 is connected with a first normal pressure discharge valve 13. The chromatographic column 7 is connected with a plurality of column position valves, wherein the plurality of column position valves include a first column position valve 19, a second column position valve 20, a third column position valve 21 and a fourth column position valve 22, two column position valves among the valve first column position valve 19, the second column position valve 20, the third column position valve 21 and the fourth column position valve 22 are combined to control forward flow, reverse flow or bypass flow through the chromatographic column 7, the input end of the chromatographic column 7 is connected with a mixer 28, a pressure sensor 29, a first bubble trap 6 and a first sensor group 31 in sequence, the first sensor group 31 includes but is not limited to an air flow meter, a mass flow meter and a pressure sensor, the output end of the chromatographic column 7 is controlled to discharge liquid by a seventh pneumatic diaphragm 23, and is connected with a liquid outlet valve group 25 and an ICE analysis module 8 (the ICE analysis module 8 is a capillary isoelectric focusing electrophoresis system, the ICE analysis module 8 is composed of a capillary, a high-voltage power supply, an isoelectric focusing electrophoresis buffer and a sample loading module, etc., by replacing the capillary isoelectric focusing electrophoresis sample loading module, the working flow can be changed, the working range and application scene of the device are maximized) through a sanitary two-position three-way reversing valve 24 respectively, and the chromatographic column 7 and the ICE analysis module 8 are further connected with a second sensor group 32, and the second sensor group 32 includes but is not limited to a pressure sensor, a conductivity sensor, a pH sensor and an ultraviolet sensor.

[0021] Preferably, the first bubble trap 6 is connected with the main pipeline 1 through a fourth pneumatic diaphragm valve 15, and the first bubble trap 6 is provided with a liquid inlet pipe, a liquid outlet pipe and an exhaust pipe, the liquid inlet pipe is provided with a third pneumatic diaphragm valve 14, the liquid outlet pipe is provided with a fifth pneumatic diaphragm valve 16, and the exhaust pipe is provided with a sixth pneumatic diaphragm valve 17. The third pneumatic diaphragm valve 14, the fourth pneumatic diaphragm valve 15 and the fifth pneumatic diaphragm valve 16 are used to control the protein chromatography purified sample to enter the first bubble trap 6.

[0022] Preferably, the outer side wall of the first bubble trap 6 is provided with a first liquid level sensor 30, which can detect the gas in the first bubble trap 6, and when the gas is excessive, the excessive gas can be discharged to avoid the gas entering the chromatography column 7; the first bubble trap 6 is also connected with a second normal pressure discharge valve 18, which can be used for balancing the internal and external pressures, and the second normal pressure discharge valve 18 has simple structure and good exhaust effect.

[0023] In addition, as shown in the above structure: Figure 2 The chromatography column 7 includes a column body 71 and the second bubble trap 72, the liquid inlet end of the column body 71 is provided with the second bubble trap 72, the outer side wall of the second bubble trap 72 is provided with a second liquid level sensor 73, and the second bubble trap 72 is provided with a liquid inlet port 74 for connecting the chromatography system, a liquid outlet port 75 for connecting the liquid inlet end of the column body 71, and an exhaust port 76 for discharging the gas captured by the bubble trap, and each port is provided with a valve. The second bubble trap 72 is additionally arranged on the chromatography column 7, which is used for removing the bubbles generated in a section of the main pipeline 1 between the first bubble trap 6 and the chromatography column 7, and also can be used for removing the bubbles for the second time, thereby improving the defoaming efficiency, and thus helping to improve the protein chromatography purification quality.

[0024] Specifically, in the structure of the chromatography column 7, the inside of the second bubble trap 72 is provided with a defoaming area 721 and a gas-liquid mixing area 722, the defoaming area 721 is the adjacent area of the liquid inlet port 74, and the gas-liquid mixing area 722 is the adjacent area of the exhaust port 76.

[0025] More specifically, the cross-sectional area of the defoaming area 721 is smaller than that of the gas-liquid mixing area 722. The volume difference between the defoaming area 721 and the gas-liquid mixing area 722 not only can reduce the liquid flow rate, but also can quickly break the bubbles through the instantaneous volume change, thereby improving the defoaming efficiency.

[0026] In the technical scheme of the utility model, the combination of the chromatography structure and the ICE analysis structure is the technical key of the case, Figure 1 The key points of the case are the related components and specific structures involved in the chromatography structure and the ICE analysis structure, and two groups of bubble traps are arranged, which can fully discharge the bubbles and improve the protein chromatography purification quality and analysis accuracy. For the pump body, the valve and the sensor, the ordinary skilled in the art can make routine settings according to the prior art, and the case has no special requirements for the model selection and combination use.

[0027] Thus, by adopting the technical scheme of the protein chromatography purification sample is transmitted to the ICE analysis module 8 for detection and analysis after the chromatography column 7 chromatography purification and a series of detection sensors detect that the standard is reached, the protein chromatography purification sample is transmitted to the ICE analysis module 8 for detection and analysis, on the basis of completing the biopharmaceutical purification, synchronous rapid protein qualitative detection is carried out, the detection efficiency is effectively improved, time is saved for subsequent protein product processing, and the protein quality is further improved under the premise of reserving the activity of the biopharmaceutical product.

[0028] It can be found from the above description that, compared with the prior art, after the technical scheme of the present application is adopted, protein full-automatic chromatography is carried out through chromatography flow path combination, online real-time protein chromatography purification operation is completed, the ICE (capillary isoelectric focusing electrophoresis system) analysis module is switched through the sanitary two-position three-way directional valve, the sterile environment of chromatography production can be effectively avoided from being damaged, the ICE (capillary isoelectric focusing electrophoresis detection) is used instead of the traditional gel plate isoelectric focusing detection, online real-time protein qualitative analysis can be carried out simultaneously, the analysis speed is improved, various application scenarios from small-scale development to pilot amplification can be met, one machine is used for multiple purposes; moreover, the detection time can be greatly shortened by using the ICE analysis module to detect protein online, the product property results are analyzed online simultaneously in the collection time period of protein chromatography purification, the product activity is maximized, and more reliable quality guarantee is provided for biopharmaceutical production.

[0029] The technical scheme, working process and implementation effect of the present application are described in detail above, and it should be noted that the described is only a typical example of the present application, in addition to this, the present application can have other various specific implementation manners, and any technical scheme formed by equivalent replacement or equivalent transformation falls within the scope of the present application.

Claims

1. A protein chromatography purification and detection integrated device, comprising a first pump body (4), a second pump body (5), a chromatography column (7) and an ICE analysis module (8), the first pump body (4) is arranged on a first branch pipeline (2), the second pump body (5) is arranged on a second branch pipeline (3), the chromatography column (7) and the ICE analysis module (8) are arranged on a main pipeline (1), characterized in that: The output end of the first pump body (4) is converged to the main pipeline (1) through the first branch pipeline (2), the input end of the first pump body (4) is connected with an air flow meter (26) and a first liquid inlet valve group (9) in sequence, and the output end of the first pump body (4) is connected with a first pneumatic diaphragm valve (10); the first output end of the second pump body (5) is converged to the main pipeline (1) through the second branch pipeline (3), the input end of the second pump body (5) is connected with a second liquid inlet valve group (11), the first output end of the second pump body (5) is connected with a mass flow meter (27) and a second pneumatic diaphragm valve (12) in sequence, and the second output end of the second pump body (5) is connected with a first normal pressure discharge valve (13); the chromatographic column (7) is connected with a plurality of column valves, the input end of the chromatographic column (7) is connected with a mixer (28), a pressure sensor (29), a first bubble trap (6) and a first sensor group (31) in sequence, and the output end of the chromatographic column (7) is controlled to discharge liquid by a seventh pneumatic diaphragm (23) and connected with a liquid outlet valve group (25) and an ICE analysis module (8) through a sanitary two-position three-way reversing valve (24) respectively. ​ 2. The protein chromatography purification and detection integrated device according to claim 1, wherein: The first bubble trap (6) is connected with the main pipeline (1) through a fourth pneumatic diaphragm valve (15), and the first bubble trap (6) is provided with a liquid inlet pipe, a liquid outlet pipe and an exhaust pipe, the liquid inlet pipe is provided with a third pneumatic diaphragm valve (14), the liquid outlet pipe is provided with a fifth pneumatic diaphragm valve (16), and the exhaust pipe is provided with a sixth pneumatic diaphragm valve (17).

3. The protein chromatography purification and detection integrated device according to claim 2, wherein: The outer side wall of the first bubble trap (6) is provided with a first liquid level sensor (30).

4. The protein chromatography purification and detection integrated device of claim 2, wherein: The first bubble trap (6) is further connected with a second normal pressure discharge valve (18).

5. The protein chromatography purification and detection integrated device of claim 1, wherein: The chromatographic column (7) and the ICE analysis module (8) are further connected with a second sensor group (32).

6. The protein purification and detection integrated device according to claim 1 or 5, wherein: The chromatographic column (7) comprises a column body (71) and a second bubble trap (72), the liquid inlet end of the column body (71) is provided with the second bubble trap (72), the outer side wall of the second bubble trap (72) is provided with a second liquid level sensor (73), and the second bubble trap (72) is provided with a liquid inlet port (74) for connecting the chromatographic system, a liquid outlet port (75) for connecting the liquid inlet end of the column body (71) and an exhaust port (76) for discharging the gas captured by the bubble trap.

7. The protein chromatography purification and detection integrated device of claim 6, wherein: The inside of the second bubble trap (72) is provided with a defoaming area (721) and a gas-liquid mixing area (722), the defoaming area (721) is an adjacent area of the liquid inlet port (74), and the gas-liquid mixing area (722) is an adjacent area of the exhaust port (76).

8. The protein chromatography purification and detection integrated device of claim 7, wherein: The cross-sectional area of the defoaming area (721) is smaller than that of the gas-liquid mixing area (722).

Images