Synthesis Of Covalent Organic Framework Materials And Its Application In Enrichment Of Glycopeptides

Protein is the main undertaker of biological life activities,and the realization of its function involves several post-translational modification processes,the most important being glycosylation and phosphorylation.Among them,glycosylation modification of protein is one of the most common post-translational modification processes.The development of biological mass spectrometry technology has enabled people to effectively detect glycoproteins or glycopeptides at the molecular level.However,compared with other proteins,the number of glycoproteins is small,and the abundance of glycopeptides after enzymatic hydrolysis is low,so some Means to effectively enrich glycopeptides to improve their detection abundance,the development of new enrichment materials is the direction and focus of the field of glycopeptide enrichment.In this paper,a new type of covalent organic framework material(T-D-COF)and its oxidation product(O-T-D-COF)were prepared.After the material was characterized,it was applied to the selective enrichment of glycopeptides,and the enrichment The conditions were optimized,and the most suitable conditions for glycopeptide enrichment were found,which laid the foundation for in-depth research.Through experiments,the detection limit of O-T-D-COF enrichment is 5 fmol/μL,the enrichment capacity is 120 mg/g,and the enrichment recoveries are 103.5 and 101.5,indicating that O-T-D-COF has a certain application in the field of glycopeptide enrichment.Prospects:Two types of COFs materials have been synthesized by hydrothermal synthesis:COF-5 and COF-10.Due to the restriction of material synthesis conditions,the crystal structure of the material is lacking.The remaining hydroxyl and boron hydroxyl groups can effectively enrich glycopeptides and significantly improve the enrichment selectivity of glycopeptides.The selectivity of glycopeptide enrichment of the two materials in the complex sample system was further verified.At the same time,COF-5 was post-modified,and a new type of composite material(COF-5-DA-Carr)was synthesized.The glycopeptide enrichment properties of the materials are compared and the reasons are analyzed,which will provide a reference for the development of modified phases of glycopeptide enrichment materials in the future;a new type of covalent organic framework material(HB-TP)has been designed and synthesized.After system characterization,its effect on glycopeptide enrichment was investigated.HB-TP is used as the matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS)detection matrix for glycopeptide detection.Compared with the traditional matrix(DHB/phosphoric acid),the effect of HB-TP itself as a matrix is the same as that of DHB/phosphoric acid The matrix is comparable.This preliminarily shows that HB-TP can be used as a glycopeptide enrichment material(enriched to 29 glycopeptide peaks)and a biological mass spectrometry detection matrix(with a strong signal peak intensity)at the same time,providing for the development of glycoproteomics experimental methods A new idea.In summary,this work focuses on glycopeptide enrichment,synthesizes several COF materials,explores the reasons that affect the glycopeptide enrichment performance of the materials from many aspects,and explores the possibility of COFs materials in the field of glycopeptide enrichment.Providing a new idea for the development of experimental methods of glycoproteomics.