Preparation Of Novel Phenyl Boronic Acid Functionalized Material By Thiol-Yne Click Chemistry And Application In Glycoprotein/Glycopeptide Selective Enrichment

The application of boronic acid affinity chromatography in glycoprotein/glycopeptide enrichment tends to be more and more mature. The enrichment selectivity , biocompatibility , and facile operation protocol are key points for efficient enrichment methods. In this work, a novel boronic acid functionalized material (TYBA material) was prepared by using Thiol-yne click chemistry. The TYBA material was proved to be successfully synthesized by infrared (IR) characterization. The glycopeptide/glycoprotein enrichment selectivity of the TYBA material was evaluated. Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied for evaluation of glycopeptide enrichment selectivity. Taking digest of horseradish peroxidase (HRP) and immunoglobulin G (IgG) as samples , 11 and 8 glycopeptides were identified with improved signals after TYBA material enrichment, respectively. High abundant non-glycopeptides were well removed from the eluting fraction. The result indicated high glycopeptide enrichment selectivity of TYBA material. A mixture of HRP and bovine serum albumin (BSA) enzymatic solution (molar ratio 1:10) was used as the sample and 8 glycopeptide signals were identified after enrichment. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied for evaluation d of enrichment selectivity of glycoprotein. The mixture of HRP, IgG, BSA and RNaseB proteins was used as model sample, and the results showed that TYBA material had high glycoprotein enrichment selectivity. The application of TYBA material in the analysis of real biosample was also evaluated by human plasma. The results indicated the TYBA material could be used in large scale glycoprotein analysis.