A New Method For Identification Of High - Throughput Glycated Peptides Based On Mass Spectrometry

This assay describes a new mass-spectrometry based method for high-throughput glycopeptide analysis. The main innovations of this methods are:1) by incorporating data-dependent database and network-centric algorithm, the efficiency of glycopeptide spectra elucidation is significantly improved; 2) alternative CID/HCD spectra collection is used as the platform for software development. The contribution of this assay is:the author developed automated software for high-throughput glycopeptide spectrum identification, and discovered more than 700 glycopeptides in human serum sample, which is the best result for software aimed at glycosylation. This assay is divided into two parts: background (chapter 1,2) and research work (chapter 3-6).The first two chapters describe the leading edge of glycopeptide analysis. In the first chapter, the current situation of post-translational modification based proteomics and glycobiology is introduced, and cross field of the two is glycoproteomics. In the second chapter, complexity of glycoproteins and mass-spectrometry based method are systematically analyzed, and current methods for glycopeptide analysis are presented.Chapter three to six are research progress and result demonstration for the software. Chapter three describes the initial phase of software development, which includes manual annotation of thousands of spectra and the realization of automation. Chapter four focuses on quality control and result validation. Possible solutions are discussed and alternative CID/HCD spectra collection is the main solution. Chapter five is the further optimization of algorithm and demonstration of the final result. Finally, chapter six lists high-quality glycopeptide spectra, result of high-throughput glycosylation analysis on some importance proteins, and the experimental procedures.