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Affinity chromatography medium as well as preparation method and application thereof

A chromatographic medium and affinity technology, applied in the field of protein separation and purification, can solve the problem of low ligand loading, and achieve the effects of improving solid loading, efficient affinity chromatography process and stable structure

Active Publication Date: 2021-01-12
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This patent retains the feature of high selectivity of peptide ligands for antibodies, and the elution conditions are mild, but there are still cases where the ligand loading is low

Method used

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  • Affinity chromatography medium as well as preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] An affinity chromatography medium, the preparation method is as follows:

[0065] (1) Preparation of ultra-macroporous matrix

[0066] Referring to CN105037626A, the ultra-large pore microspheres are prepared, and the average pore diameter is 102.4nm.

[0067] (2) The epoxy groups carried by the ultra-macroporous matrix are reduced to hydroxyl groups

[0068] Accurately weigh 1.0 g of ultra-large pore microspheres into a 50 mL Erlenmeyer flask, and then add 2 M H 2 SO 4 The aqueous solution was incubated at 40° C. in a 120 rpm vibrating shaker for 2 hours, then suction-filtered, washed with deionized water, and suction-filtered until there was no obvious water on the surface of the microspheres after washing.

[0069] (3) Preparation of polymer brushes on the substrate surface

[0070] Put the super-macroporous microspheres reduced to hydroxyl groups into a cerium ammonium nitrate solution with a pH of 9.0 and a concentration of 3.0%, react at 65°C for 0.5h, add 2mL...

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Abstract

The invention provides an affinity chromatography medium as well as a preparation method and application thereof. The affinity chromatography medium comprises a super-macroporous matrix, a polymer brush and a polypeptide ligand, wherein the polypeptide ligand is connected with the macroporous matrix through the polymer brush. The affinity chromatography antibody provided by the invention has the advantages of stable structure, salt resistance, acid and alkali resistance, difficult ligand shedding, easy regeneration and the like, and the purity of the antibody purified by the affinity chromatography medium provided by the invention can reach 94% or above.

Description

technical field [0001] The invention belongs to the technical field of protein separation and purification, and relates to an affinity chromatography medium and its preparation method and application. Background technique [0002] Monoclonal antibodies have been widely used in the fields of clinical medicine and biotechnology because of their specificity and high purity, and have rapidly promoted the development of downstream purification processes for monoclonal antibodies. At present, the downstream purification of monoclonal antibody drug industry mainly uses affinity chromatography ligands based on Protein A and its derivative proteins. Recombination of these protein ligands has excellent specificity and selectivity for antibodies, and is widely used in the separation and purification of antibodies, but there are still defects such as expensive, easy to fall off, and acid and alkali resistance, which become the main material cost in the antibody production process . In...

Claims

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Application Information

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IPC IPC(8): B01J20/286B01J20/30B01D15/38C07K1/22C07K16/00
CPCB01J20/286B01D15/3809C07K16/00C07K1/22
Inventor 马光辉郝冬霞张荣月葛佳黄永东赵岚周炜清
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI