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Three-dimensional liquid chromatography protein purification device and using method thereof

A protein purification, three-dimensional liquid phase technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of cumbersome, time-consuming and labor-intensive multiple offline purification processes, and achieve convenient operation, less time-consuming, and good reproducibility. Effect

Active Publication Date: 2018-12-18
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a three-dimensional liquid chromatography protein purification device and its use method to solve the problems of cumbersome, time-consuming and labor-intensive multiple off-line purification processes in the current protein chromatography purification process

Method used

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  • Three-dimensional liquid chromatography protein purification device and using method thereof
  • Three-dimensional liquid chromatography protein purification device and using method thereof
  • Three-dimensional liquid chromatography protein purification device and using method thereof

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Embodiment 1

[0047] The first chromatographic column Z1 is a phenyl sepharose hydrophobic interaction chromatography column with a size of 310*26mm; the second chromatographic column Z2 is a diethylaminoethyl (DEAE) dextran gel weak anion exchange column with a size of 310*26mm ; The third chromatographic column Z3 is a dextran and epichlorohydrin cross-linked molecular exclusion chromatography column with a size of 310*26mm. On-line multi-stage protein purification device such as figure 1 As shown, there are three sets of pump systems including the first pump system P1 (phase A and phase B are P1A and P1B respectively), the second pump system P2 (phase A and phase B are P2A and P2B respectively), and the third pump system P3; The first switching valve V1 and the second switching valve V2, wherein the switching valves each have 6 connection points of numbers 1-6, and have two working positions of A and B. Wherein the first pump system P1 is connected with the first chromatographic column ...

Embodiment 2

[0060] The first chromatographic column Z1 is a butyl dextran gel hydrophobic interaction chromatography column, specifications

[0061] 310*26mm; the second chromatographic column Z2 is a carboxymethyl sepharose weak cation exchange column with a specification of 310*26mm; the third chromatographic column Z3 is allyl dextran and N,N-methylenebisacrylamide Cross-linked molecular exclusion chromatography column, size 310*26mm. The device connection mode is as in Example 1, the mobile phase of the first chromatographic column Z1 is phosphate buffer, the mobile phase of the second chromatographic column Z2 is ammonium formate buffer salt, and the mobile phase of the third chromatographic column Z3 is pure water. Initially, the first switching valve V1-is placed at position A, and the impurity protein before the first chromatographic column Z1 directly flows into the waste liquid pool. When the target protein starts to elute, switch the first switching valve V1 to position B, at ...

Embodiment 3

[0064] The first chromatographic column Z1 is an antigen polypeptide column with a size of 310*26mm; the second chromatographic column Z2 is a carboxymethylstyrene-divinylbenzene copolymer weak cation exchange column with a size of 310*26mm; the third chromatographic column Z3 is a high Cross-linked agarose molecular exclusion chromatography column, size 310*26mm.

[0065] The device connection method is as in Example 1. The mobile phase of the first chromatographic column Z1 is Tris-HCl salt buffer, the mobile phase of the second chromatographic column Z2 is ammonium acetate buffer salt, and the mobile phase of the third chromatographic column Z3 is pure water. The operating principle and steps are as in Example 1.

[0066] Those of ordinary skill in the art should be able to understand that one of the characteristics or purposes of the present invention is that the three-dimensional liquid chromatography protein purification device and its use method proposed by the present ...

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Abstract

The invention provides a three-dimensional liquid chromatography protein purification device and a using method thereof. The device comprises three sets of infusion pump systems, three chromatographiccolumns and two switching valves. The chromatographic columns of three different retention mechanism are combined on line, and the switching valves are controlled, so that target protein is selectively purified step by step through the three chromatographic columns, and thus the high-purity protein is prepared. The chromatographic columns of the three retention mechanisms are integrated, so thatcontinuous operation can be realized, the automation degree is high, and the reproducibility is good. One-time sampling is realized, and the high-purity target protein is purified in one step. Compared with a traditional off-line separation mode, the loss of a sample during fractionated separate collection, enrichment, re-sampling and other treating processes can be avoided, and the risk of denaturation, pollution and the like of the sample in the treating processes is reduced, and the time and labor cost of step-by-step treatment are greatly saved.

Description

technical field [0001] The invention belongs to the field of biomedical purification devices, and in particular relates to a three-dimensional liquid chromatography protein purification device and a use method thereof. Background technique [0002] Drugs such as genetically engineered drugs, monoclonal antibodies, and recombinant vaccines are all proteins in nature. Compared with traditional small molecule drugs, biological drugs have the advantages of strong activity, strong specificity, and clear biological functions. The protein extraction process is difficult to control, and the low purification yield is the bottleneck of biopharmaceutical development. According to reports, the downstream process cost of biopharmaceuticals accounts for 80% of the drug production cost, which shows that the purification process has a great impact on the promotion of biopharmaceuticals. In recent years, with the rapid development of the biomedical industry, the bottleneck of protein purifi...

Claims

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Application Information

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IPC IPC(8): G01N30/20
CPCG01N30/20
Inventor 李笃信易琳朱梦张真庆邱飘飘闫娜
Owner SUZHOU UNIV