Pretreatment composition, pretreatment method and application of abnormal protein of sugar chain structure suitable for mass spectrometric detection

Abstract

The invention relates to a pretreatment composition, a pretreatment method and application of a sugar chain structure abnormal protein suitable for mass spectrometric detection. According to the pretreatment method for the protein with the abnormal sugar chain structure, the protein with the abnormal sugar chain structure in a sample can be accurately separated; the enriched protein with the abnormal sugar chain structure does not need other treatment, and the mass spectrometric detection can be directly carried out; a simple pretreatment method is provided for mass spectrometric detection andanalysis, so a mass spectrometric analyzer can screen and detect related diseases such as tumors and autoimmune diseases. According to the invention, the method has important significance in mass spectrometric detection and extremely high sensitivity; the method is rapid, simple and convenient, and the method can be carried out without magnetic separation equipment or automatic equipment.

Description

technical field [0001] The invention belongs to the technical field of medical equipment and in vitro diagnosis, and relates to a pretreatment composition, a pretreatment method and an application of a protein with an abnormal sugar chain structure suitable for mass spectrometry detection. Background technique [0002] The basic principle of mass spectrometry (MS) is to ionize particles such as molecules or atoms in the ion source, and under the action of an accelerating electric field, let the molecules or atoms of the ionized pair enter the mass analyzer. 1. The magnetic field causes various ions to disperse and focus according to the mass-to-charge ratio, so as to determine their mass, and determine the type and quantity of the sample to be tested according to the mass and spectrum analysis. [0003] Since the 1940s, mass spectrometry has been widely used in the analysis of organic substances, and gas chromatography-mass spectrometry (GC-MS) technology, field desorption (...

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Problems solved by technology

[0010] The above-mentioned pretreatment methods for glycosylated protein spectrum detection must go through multi-step processes such as enzyme digestion, sugar chain dissociation, and desalination, before mass spectrometry detection and analysis of glycosylated protein sugar chains or glycopeptides can be performed. The operation is complex and Expensive, which greatly limits the application as a pretreatment method for mass spectrometry

[0011] Moreover, after the above pretreatment methods, mass spectrometry does not directly detect macromolecular proteins such as glycosylated proteins, such as directly detecting proteins with abnormal sugar chain structures with a relative molecular weight greater than 10,000 Daltons (10KDa), but detects glycosylated proteins mass spectrometry detection and analysis of sugar chains or glycopeptides, but the detection of sugar chains has a relatively restrictive effect on the diagnosis of diseases and is not suitable for direct use in the diagnosis of related diseases

It has been reported that many different types of sugar chains can be identified by releasing sugar chains and performing mass spectrometry detection; however, the occurrence of diseases is often related to abnormal glycosylation of specific proteins, and sugar chain analysis alone cannot accurately identify whether it is a specific protein. Abnormally glycosylated sugar chains, which have a great limitation on the diagnostic application of diseases

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