Boronic acid modified magnetic nanoparticles for enrichment of glycoproteins via azide and alkyne click chemistry

Abstract

In this study, a novel approach was developed to synthesize aminophenylboronic acid functionalized magnetic nanoparticles (NPs) via Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) 'click' chemistry. Firstly, azide-functional Fe 3 O 4 NPs were obtained by a two-step chemical modification process. Then, an alkyne-phenylboronic acid molecule was connected onto the surface of magnetite by the CuAAC reaction. The morphology, structure and composition of the synthesized nanocomposites were characterized by transmission electron microscopy (TEM), X-ray powder diffraction (XRD), vibrating sample magnetometer (VSM), Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectrometry (XPS). Five proteins, including ovalbumin (OB), transferrin (Trf), as glycoprotein templates and lysozyme (Lyz), bovine serum albumin (BSA), horse heart cytochrome c (Cyt C) as nonglycoprotein templates are chosen as target proteins. The as-prepared clickFe 3 O 4 @APBA NPs with a mean diameter of 23.2 nm showed a strong magnetic response to an externally applied magnetic field and exhibited a high adsorption capacity and excellent specificity towards glycoproteins in comparison with nonglycoproteins. The click-Fe 3 O 4 @APBA NPs showed the higher adsorption capacity towards glycoproteins than the nonclick-Fe 3 O 4 @APBA NPs which were synthesized through a common nucleophilic substitution reaction. The greatly enhanced adsorption capacity towards glycoproteins demonstrated that the 'click' method presented great superiority in ligand immobilization. Finally, the click-Fe 3 O 4 @APBA NPs could efficiently enrich glycoproteins from real egg white samples as well.

Synthesis of boronate-silica hybrid affinity monolith via a one-pot process for specific capture of glycoproteins at neutral conditions

“…In addition, the moderate acidic elution condition make boronate affinity well coupled with mass spectrometry for analysis of cis-diol biomolecules in -omics [14][15][16]. Different types of boronate affinity materials have been developed for specific capture of glycoproteins [15][16][17][18][19][20].…”

Global and site‐specific analysis of protein glycosylation in complex biological systems with Mass Spectrometry

“…The covalent interactions between boronic acids (BA) and cis ‐diols on glycans have been extensively studied and been applied for glycoprotein analysis (Dowlut and Hall, ; Jin et al, ; Zhang et al, ; Wang et al, ; Xu et al, ; Wang et al, ; Wu et al, ; Xiao et al, ; Liu and He, ; Xiao et al, ). Yang and co‐workers designed a boronic acid‐functionalized core‐satellite‐structured composite material to capture glycopeptides/proteins, and identified 194 unique glycosylation sites from 155 different glycoproteins (Zhang et al, ).…”

Chemistry in Separation and Enrichment of Glycoproteins/Glycopeptides

“…张玉奎研究组制备的硼酸修饰 的纳米金粒子能从 HRP/BSA 的混合蛋白中高选择性地 富集 HRP, 该硼酸修饰的纳米粒成功规模化地鉴定识别 了人类血浆糖蛋白质的 160 个糖蛋白 [24] . 该研究组还制 备了不同含硼酸配体修饰的磁性粒子, 并研究了粒子对 糖蛋白进行分离富集, 磁性粒子在 pH＝7.4 的生理条件 下能对鸡蛋清中的糖蛋白进行分离富集 [25,26] . 目前用于 …”