Glycoprotein N-sugar chain analysis method

Abstract

The invention relates to the technical field of biology, in particular to a glycoprotein N-sugar chain analysis method. The method comprises the following steps: a) carrying out protein expansion denaturation and disulfide bond reduction treatment on glycoprotein to be analyzed by using 2-methylpyrimidine borane and a disulfide bond reducing agent; b) carrying out alkylation treatment on the protein treated in the step a), and carrying out enzyme digestion on N-glycosidase F; and c) drying the sample subjected to enzyme digestion, marking the dried sample with a marker, and analyzing the N-carbohydrate chain according to the marker, wherein the marker is procaine amide and / or a salt thereof. The method can obviously shorten the operation process and increase the detection flux, and is more environment-friendly and easy to use.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for analyzing glycoprotein N-sugar chains. Background technique

[0002] During the production of recombinant proteins, conditions such as culture conditions will affect the level of post-translational modification of cellular proteins. Glycosylation is one of the most common post-translational modifications of proteins in eukaryotic cells. It plays a vital role, such as protein folding, cell recognition, cancer cell metastasis, membrane protection, blood group determination system, material transport, etc.

[0003] The most common glycoproteins in organisms are N-glycosidic bonded glycoproteins, that is, proteins modified by linking oligosaccharides to the amide nitrogen of the asparagine side chain on the peptide chain. N-glycan chains only account for a small part of the entire glycoprotein, but have a huge impact on the properties of the entire glycoprotein, so N-glycan...

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