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Biomimetic affinity purification method of para-aminophenamidine biomimetic affinity ligand

A technology of p-aminobenzamidine and purification method, which is applied in the field of biomimetic affinity purification of p-aminobenzamidine biomimetic affinity ligands, can solve the problems of increasing steps and costs, affecting the utilization of metalloprotease, etc., and achieves a simple process. Effect

Active Publication Date: 2018-06-12
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, in the process of metal chelation chromatography, imidazole with certain toxicity needs to be used in the elution process, which will affect the utilization of metalloproteases as pharmaceuticals, and the use of desalting columns to process imidazole will increase additional steps and costs.

Method used

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Embodiment 1

[0035] 1. Synthesis of biomimetic affinity materials

[0036] 1), epichlorohydrin activated agarose gel

[0037] The agarose gel (Sepharose 6B) was thoroughly washed with double distilled water at a ratio of 1:10 V / V until the pH of the solution reached 7.0, and the washed agarose gel was air-dried and then dissolved in the activation solution (1M Sodium hydroxide, 2.5g dimethyl sulfoxide and 10ml epichlorohydrin), shake at 100rpm in a shaker at 40°C for 2.5h for activation to obtain epichlorohydrin-activated sepharose 6B (see process a).

[0038] 2), amination of epichlorohydrin activated agarose gel

[0039] Add 35% saturated ammonia to the activated agarose gel 6B, then add a certain amount of double distilled water, and incubate overnight at 100 rpm in a shaker at 30°C, so that the activated agarose gel 6B is added with amino groups to obtain aminated activated agarose gel. Glue 6B (see procedure b).

[0040] 3), connecting the cyanuric chloride linking arm

[0041] Ad...

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Abstract

The invention relates to a biomimetic affinity purification method of a para-aminophenamidine biomimetic affinity ligand. The biomimetic affinity purification method of the para-aminophenamidine biomimetic affinity ligand comprises preparation of a biomimetic affinity material as well as biomimetic affinity purification on metalloprotease (MP) by a para-aminophenamidine agarose gel column. The purification method is simple in flow, short in cycle and high in efficiency, and the purity is up to 98.7 percent.

Description

technical field [0001] The invention relates to the technical field of marine microorganisms, in particular to a biomimetic affinity purification method of p-aminobenzamidine biomimetic affinity ligand. Background technique [0002] The biomimetic affinity ligands of marine microbial enzymes are usually derived from specific inhibitors of the enzymes. In previous studies, based on the crystal structure information of marine low-temperature alkaline protease MP, it was prepared by fermentation of marine bacteria YS-80 (marine bacteria YS-80 was isolated from deep sea mud marine bacteria (YS-80), and was preserved in CCTCC, Wuhan, China , the preservation number is CCTCC NO:M209177, see patent application number 200910177301.0, a small molecule inhibitor library of marine low-temperature alkaline protease MP was constructed through virtual screening. Among the 15 most effective small molecule reversible inhibitors predicted by fine screening Five of them are benzamidine deriv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22B01J20/30B01D15/08C12N9/50
CPCB01D15/08B01J20/22C12N9/50
Inventor 李尚勇孙谧郝建华刘均忠王跃军马子宾
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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