Sample Pre-Processing for Mass Spectrometric Analysis of Glycoprotein Structure
糖蛋白结构质谱解析的样品前处理

A novel,simple and fast method based on a quadrupole ion trap(QIT) and a time-of-flight mass spectrograph was developed for the glycosylation site and glycan structure analysis of glycoprotein by in-solution digestion and in-gel digestion which did not need to enrich and label the glycopeptide.Horseradish peroxidase(HRP) and RNase B were utilised to optimize the conditions of sample pre-processing.The effects of the conditions were investigated separately and the optimal conditions were:heat denaturation of RNase B,chemical denaturation of HRP,RNase B digestion with endoproteinase Lys-C,HRP digestion with trypsin,12-16 h of digestion,50% acetonitrile-5% trifluoroacetic acid of the extraction solution,sandwich spotting method of the sample.