基于pH响应涂层柱的毛细管电泳法在线富集和分离糖蛋白
On-line enrichment and separation of glycoprotein by capillary electrophoresis based on pH response coating column
摘要
Abstract
A capillary column coated with 3-aminophenylboronic acid(APBA)-functionalized gold nanoparticles(AuNPs@APBA)was prepared via electrostatic self-assembly.The coated column exhibited anti-nonspecific adsorption of glycoproteins,enabling selective online enrich-ment during capillary electrophoresis(CE).First,gold nanoparticles(AuNPs)were synthe-sized using the sodium citrate reduction method.Then,APBA was self-assembled electrostati-cally on the surface of the AuNPs to obtain AuNPs@APBA.This nanomaterial was bonded to the inner wall of a capillary through ion adsorption to produce a AuNPs@APBA-coated capillary column.Glycoproteins were adsorbed via bond formation with boric acid groups under alkaline conditions(pH 8)to generate borate esters.Under acidic conditions(pH 3),the borate esters dissociated to release the glycoproteins,thereby achieving the selective online enrichment and separation of glycoproteins.The AuNPs and AuNPs@APBA were characterized using Fourier transform infrared spectroscopy,and their sizes and Zeta potentials were determined.In addition,the electroosmotic flow(EOF)of the AuNPs@APBA-coated capillary column was measured.The results showed that the surface of the AuNPs was successfully modified with APBA and that AuNPs@APBA was adsorbed on the inner wall of the capillary.The peak area of ovalbumin(OVA)on the AuNPs@APBA-coated column was 26.46 times higher than that on a bare column via conventional electrophoresis.In contrast,the peak area of bovine serum albu-min(BSA)only increased by 8.47 times,indicating that the AuNPs@APBA coated column se-lectively enriched glycoproteins.Evaluation of the reproducibility and stability of this method revealed that the AuNPs@APBA coated capillary column could be used continually for 33-67 h.The relative standard deviations(RSDs)of the peak areas for intra-day(n=5)and inter-day(n=6)analyses were 2.2%and 3.0%,respectively.The developed method was successfully applied to enrich glycoproteins in a 1×106-fold diluted egg white sample.Glycoproteins were not detected using conventional electrophoresis on the bare column,whereas the AuNPs@APBA-coated capillary column effectively enriched and separated glycoproteins,resulting in a peak area of 10 469 mAU·ms.Furthermore,the entire enrichment and separation process was com-pleted within 3 min.This new online enrichment and separation method for glycoproteins has the advantages of low sample consumption,simple operation,and high separation efficiency.关键词
毛细管电泳/在线富集/离子吸附/糖蛋白/苯硼酸Key words
capillary electrophoresis(CE)/on-line enrichment/ion adsorption/glycoprotein/phenylboronic acid分类
化学化工引用本文复制引用
张剑,徐彩凤,马彩莲,贺茂芳,张博,刘春叶..基于pH响应涂层柱的毛细管电泳法在线富集和分离糖蛋白[J].色谱,2024,42(9):903-908,6.基金项目
陕西省科学技术厅社会发展项目(2023-YBSF-202) （2023-YBSF-202）
西安医学院大学生开放实验科研基金项目(2021DXS50,2021DXS68) （2021DXS50,2021DXS68）
西安医学院秦创原·科技成果产业化培育项目(21CYH01) （21CYH01）
西安医学院校内人才项目(2018XNRC03) （2018XNRC03）
西安医学院科研能力提升计划项目(2022NLTS063,2022NLTS069).Social Development Project of Shaanxi Provincial Department of Science and Technology (No.2023-YBSF-202) （2022NLTS063,2022NLTS069）
Open Experimental Research Fund Project for College Students of Xi'an Medical University (Nos.2021DXS50,2021DXS68) （Nos.2021DXS50,2021DXS68）
Qin Chuangyuan Science and Technology Achievement Industrialization Cultiva-tion Project of Xi'an Medical University (No.21CYH01) （No.21CYH01）
Campus Talent Project of Xi'an Medical Univer-sity (No.2018XNRC03) （No.2018XNRC03）
Research Capability Enhancement Plan Project of Xi'an Medical University (Nos.2022NLTS063,2022NLTS069). （Nos.2022NLTS063,2022NLTS069）
