System and method for solid phase enrichment coupled mass spectrometric detection of glycoprotein

Abstract

The invention discloses a system and a method for the solid phase enrichment coupled mass spectrometric detection of glycoprotein. The system comprises an acylhydrazine modified porous enzyme reactor and a mass spectrograph, wherein the acylhydrazine modified porous enzyme reactor is used for carrying out selective enrichment and quick in-situ enzymolysis on the glycoprotein by adopting an acylhydrazine treated porous silica material as an adsorbent; enzymatic hydrolysate is subjected to mass spectrometric analysis by the mass spectrograph. According to the system and the method, the selective enrichment and the quick in-situ enzymolysis of the glycoprotein are realized through utilizing the pore canal limitation effect of the porous silica material and the chemical affinity effect between an acylhydrazine modified surface and the glycoprotein; the mass spectrometric direct analysis is further completed. The system and the method are simple in step, convenient to operate, quick and high-efficiency, and can be used for realizing the high-sensitivity, high-accuracy and high-flux mass spectrometric analysis on the glycoprotein.

Description

technical field [0001] The invention belongs to the technical field of biochemical analysis, and relates to a solid-phase enrichment method for glycoproteins and mass spectrometry detection and analysis of glycoproteins. Background technique [0002] Studies have revealed that glycosylation is a post-translational modification ubiquitous in organisms and performs important biological functions. Glycosylation modification plays an important role in various life phenomena, such as participating in cell adhesion and signal transduction, affecting protein secretion and stability, immune and inflammatory responses, and affecting the direction of protein transfer in cells. Studies have reported that more than 1 / 2 of proteins in organisms are glycosylated. More importantly, some glycoproteins are generally considered important biomarkers, so their detection is often closely related to related pathological or physiological processes. For example, Her2 / neu in breast cancer, prostat...

AI Technical Summary

Problems solved by technology

First of all, although there are many types of proteins containing glycosylation modifications, their abundance is usually low, and the proportion of all peptides after enzymatic hydrolysis into peptides is even lower, usually only about 1-3% of the peptides are There are glycosylation modifications; second, the ionization efficiency of peptides with glycosylation modifications in mass spectrometry is often lower than that of non-modified peptides, so it is not easy to be identified by mass spectrometry; third, although there are many methods to achieve The enrichment of glycoproteins has been achieved, but traditional methods, such as antigen-antibody immunoreaction and Western blotting, can only study a small amount of protein alone, and it is time-consuming

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