Filter membrane assisted separation and identification method for glycoprotein full O-linked carbohydrate chain in biological sample

A biological sample and glycoprotein technology, which is applied in the direction of analyzing materials, preparation of test samples, material analysis by electromagnetic means, etc., can solve the problems of low reaction efficiency and few types of sugar chains, etc., achieving obvious effects and wide applicability Effect

Inactive Publication Date: 2012-11-07
NORTHWEST UNIV(CN)
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  • Abstract
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Problems solved by technology

[0004] The invention provides a method for membrane-assisted separation of all O-linked sugar chains of glycoproteins in biological samples to solve the problems of low reaction efficiency and few types of sugar chains that can be identified in the prior art

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  • Filter membrane assisted separation and identification method for glycoprotein full O-linked carbohydrate chain in biological sample
  • Filter membrane assisted separation and identification method for glycoprotein full O-linked carbohydrate chain in biological sample

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Embodiment Construction

[0030] The present invention utilizes the molecular sieve effect of the 8-12KD filter membrane to separate the O-sugar chain released from the glycoprotein from the protein, such as figure 1 . The molecular weight of the protein is greater than 10KD, and the O-linked sugar chains found so far are all small and the number of sugar groups is small, and its molecular weight is generally several thousand Da. The O-linked sugar chains of glycoproteins on the 10KD molecular sieve are released by β-elimination reaction, and the O-linked sugar chains flow out by washing and repeated centrifugation, and the protein remains on the filter membrane, thus realizing the O-linked sugar chains in one step separation.

[0031] Taking serum samples as an example, the method for isolating all O-linked sugar chains of glycoproteins and identifying them is realized according to the present invention. The main steps for other protein samples (such as saliva, urine, etc.) are exactly the same as t...

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Abstract

The invention provides a filter membrane assisted separation and identification method for glycoprotein full O-linked carbohydrate chain in a biological sample and solves the problems that reaction efficiency is low, and few types of carbohydrate chains are identifiable in the prior art. By molecule sieving effect of 8-12KD filter membranes, carbohydrate chains released from glycoprotein are separated from the same, O-linked carbohydrate chains of the glycoprotein on 8-12KD molecule sieves are released by beta-elimination reaction, the O-linked carbohydrate chains flow out by means of cleaning and repeated centrifuging, protein is reserved on the filter membranes, and further separation of the O-linked carbohydrate chains is achieved. The method is effective in carbohydrate chain separation, avoids byproducts brought in due to non-specific adsorption and chemical reaction, is high in reaction efficiency and has simple reaction steps.

Description

technical field [0001] The invention relates to a method for separating all O-linked sugar chains of glycoproteins in biological samples and identifying them. Background technique [0002] Carbohydrates are biomolecules that widely exist in the body and play a variety of important roles in the body. Carbohydrates mainly exist in the form of carbohydrate complexes, such as glycolipids, glycoproteins, proteoglycans, peptidoglycans, and lipopolysaccharides. Among them, protein glycosylation is widely distributed on the cell surface and extracellular matrix. The sugar chain structure on it is related to many important biological functions, mainly involving the regulation of protein conformation and stability, and the control of protein and even cell half-life. In addition, the specific binding of these sugar chain structures as ligands mediates targeted recognition of proteins, cell-cell and cell-extracellular matrix interactions. Glycoproteins in organisms generally include m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/34G01N27/64
Inventor 李铮杨刚龙马恬然王秦哲
Owner NORTHWEST UNIV(CN)
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