Method for enriching and purifying glycosylation protein

A glycosylated protein and enrichment technology, which is applied in the field of enrichment and purification of glycosylated proteins, can solve the problems of low recovery rate, complex separation and enrichment methods of glycosylated proteins, high price, etc. The effect of low cost and simple method

Active Publication Date: 2007-05-09
SHANXI LIFEGEN
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Problems solved by technology

[0007] The purpose of the present invention is to provide a method for separating, enriching and purifying glycosylated proteins based on micro-nano magnetic technology, which solves the probl...

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  • Method for enriching and purifying glycosylation protein
  • Method for enriching and purifying glycosylation protein

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Embodiment Construction

[0046]The invention connects the lectin to the functionalized nano-scale magnetic particle, such as the nano-scale magnetic particle derivatized by epoxy ethyl group and amino group, and is used for separating and enriching the glycosylated protein. Glycosylated proteins are formed by covalently linking proteins and monosaccharide chains. They are mainly used for cell recognition, and their physiological functions are mainly used for cell recognition of the immune system.

[0047] The implementation steps of the embodiment of the present invention are as follows:

[0048] 1) Immobilization of lectins

[0049] (1) Prefabrication of the oxiranyl-derivatized magnetic particles: after shaking the oxiranyl-derivatized magnetic particles, pipette 200 μl into a centrifuge tube and mark it as No. 1 centrifuge tube. Add 400 μl of coupling buffer to No. 1 centrifuge tube, shake well by hand, and separate on a magnetic separator for 3 minutes. Keep No. 1 centrifuge tube on the magnetic ...

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Abstract

This invention discloses a method for enriching and purifying glycosylated protein. The immobilization of agglutinin comprises: adding epoxy ethyl derived magnetic particles into a centrifugal tube, performing immobilization reaction, washing, sealing, washing repeatedly to immobilize agglutinin onto epoxy ethyl derived magnetic particles, adding buffer and storing. The separation and purification of glycosylated protein comprises: immobilizing prepared agglutinin onto epoxy ethyl derived magnetic particles, adding standard glycosylated protein for adequate coupling, washing repeatedly, eluting to obtain enriched and purified glycosylated protein, loading into a dialyzing bag, and desalting. This invention solves the problems of complex enriching and separation process, low recovery rate and high cost faced by background technology. This invention has such advantages as simple process, high recovery rate, low affinity chromatographic column cost, and convenient usage.

Description

technical field [0001] The invention relates to a method for linking lectin to modified magnetic particles, using the lectin immobilized on the magnetic particles to specifically bind glycosylated proteins, thereby separating, enriching and purifying them. Background technique [0002] Lectins are a class of non-antibody proteins or glycoproteins that can specifically bind to carbohydrates non-covalently, and have the functions of aggregating cells and precipitating glycans and complex sugars. Its structure contains a carbohydrate recognition domain (CRD), and therefore can specifically bind to carbohydrates, and is specific to the anomeric configuration of residues, the type of glycosidic bonds, and the structure or conformation of oligosaccharide chains. For example, concanavalin (Con A) is a lectin from the canavalin bean, which can specifically bind α-mannose (Man) inside the sugar chain or at the non-reducing end. Concanavalin can be attached to a solid support by appr...

Claims

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Application Information

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IPC IPC(8): C07K1/14C12N9/24
Inventor 李铮陈超杨刚龙崔亚丽
Owner SHANXI LIFEGEN
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