Preparation of novel phenyl boronic acid functionalized silica gel using triazo-cyanide click chemistry and its application in glycoprotein/glycopeptide selective enrichment

doi:10.3724/SP.J.1123.2018.11015

Abstract

Abstract:

The application of boronic acid affinity chromatography to glycoprotein/glycopeptide enrichment is increasingly maturing. The enrichment selectivity, biocompatibility, and facile operation protocol are key aspects in efficient enrichment methods. In this work, a novel triazo-cyanide boronic acid functionalized material (TCNBA) was prepared using triazo-cyanide click chemistry. The TCNBA was proved to be successfully synthesized through infrared ray (IR) characterization. Subsequently, the glycopeptide/glycoprotein enrichment selectivity of the TCNBA was evaluated. Matrix-assisted laser desorption/ionization time-of-flight mass (MALDI-TOF MS) was employed for the glycopeptide enrichment selectivity evaluation. Taking the digestion of horseradish peroxidase (HRP) and immunoglobulin G (IgG) as samples, 13 and 11 glycopeptides could be characterized with improved signals after TCNBA enrichment, respectively. High abundance non-glycopeptides could be removed effectively from the eluting fraction. This result indicates the high glycopeptide enrichment selectivity of TCNBA. In addition, a mixture of HRP and bovine serum albumin (BSA) enzymatic solution (1:10, amount of substance ratio) was utilized as a sample, and five glycopeptide signals could be identified following enrichment. To evaluate the glycoprotein enrichment selectivity, sodium salt-polyacrylamide gel electrophoresis (SDS-PAGE) was adopted as an evaluation method. Mixtures of HRP, IgG, BSA, and ribonuclease B (RNaseB) proteins were employed as samples, and the results demonstrated that TCNBA had a high glycoprotein enrichment selectivity. The application of TCNBA to the analysis of a real biosample was also evaluated using human plasma. The results indicated the TCNBA could be utilized in large-scale glycoprotein analysis.

Key words: boronic acid affinity chromatography, glycopeptides, glycoprotein, selective enrichment, separation

CLC Number:

O658

ZHANG Liyuan, WANG Liheng, WANG Lili, DONG Peipei, LIU Hezhen, ZHAO Yanyan. Preparation of novel phenyl boronic acid functionalized silica gel using triazo-cyanide click chemistry and its application in glycoprotein/glycopeptide selective enrichment[J]. Chinese Journal of Chromatography, 2019, 37(3): 279-286.

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