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
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[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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