色谱Issue(4)：369-375,7.DOI:10.3724/SP.J.1123.2013.12018
伴刀豆凝集素修饰磁性纳米粒子富集人血清中糖蛋白及质谱鉴定
Enrichment of glycoproteins in human serum using concanavalin A-functionalized magnetic nanoparticles and identification by mass spectrometry
摘要
Abstract
Biomedical sciences,and in particular biomarker research,demand efficient glyco-protein enrichment platforms. Herein novel magnetic nanoparticles with an average size around 135 nm in diameter were prepared for the enrichment of glycoproteins in human serum. The prepared magnetic nanoparticles possessed uniform core / shell / shell structure which was com-posed of 8 nm magnetite internal core and double layers consisting of silica and poly glycidyl methacrylate(GMA). The latter was constructed by seed polymerization. Modified by a poly-ethylene hydrophilic linker,it made the surfaces of the magnetic nanoparticles highly hydrophil-ic so as to reduce the nonspecific adsorption of proteins. We examined affinity purification of glycoprotein in diluted human serum using our prepared magnetic nanoparticles with immobi-lization of concanavalin A( MNP@ ConA). The enriched proteins were reduced,alkylated and digested with trypsin. These peptides then were separated by offline two-dimensional chroma-tography. Protein identification was realized with nano-high performance liquid chromatogra-phy-orbitrap mass spectrometry. A total of 80 proteins were identified,among them 76 proteins were found to be glycoproteins by use of bioinformatic tools. β-2-Glycoprotein 1 present in ser-um at low mass concentration around 0. 000 01 g / L was also identified. This demonstrates the capability of magnetic nanoparticle for recovering minute amounts of glycoproteins from a fluid exhibiting a dynamic concentration range more than 12 orders of magnitude. Overall,MNP@ConA has been proven to be an efficient alternative to currently available immobilization supports.关键词
液相色谱/质谱/富集/糖蛋白/凝集素/磁性纳米粒子Key words
liquid chromatography( LC)/mass spectrometry( MS)/enrichment/glycopro-tein/lectin/magnetic nanoparticle分类
化学化工引用本文复制引用
李凤,康经武..伴刀豆凝集素修饰磁性纳米粒子富集人血清中糖蛋白及质谱鉴定[J].色谱,2014,(4):369-375,7.基金项目
国家自然科学基金项目（21175146，20975109）． （）
