表面等离子共振-质谱法对相互作用的生物分子在10^-^(15)mol水平的微量鉴定  

Identification of Interacting Molecules by Biomolecular Interaction Analysis Combined with Surface Plasmon Resonance-mass Spectrometry at 10 -15 mol Level

作　　者：聂松[1] 陈平[1] 梁宋平[1] 

机构地区：[1]湖南师范大学生命科学学院,长沙410081

出　　处：《高等学校化学学报》2005年第1期68-72,共5页Chemical Journal of Chinese Universities

基　　金：国家"八六三"计划项目 (批准号 :2 0 0 1AA2 3 3 0 3 1)资助

摘　　要：利用生物传感芯片质谱法 (BIA/MS)对微球蛋白及其抗体的相互作用进行分析和鉴定 .将微球蛋白抗体偶联到芯片上 ,让微球蛋白溶液流过芯片表面 ,然后使用“三明治”结构的微再生方法把结合的微球蛋白从芯片上洗脱下来 ,再对其进行酶解及质谱鉴定 ,在 1 0 - 1 5mol水平得到了明确的结果 .Biomolecular interaction analysis mass spectrometry(BIA/MS) is a very promising method applied in proteomics for the characterization of protein-protein interactions. BIA/MS is an approach combining the surface plasmon resonance(SPR) technology with mass spectrometry for the real-time analysis and identification of interaction molecules that interact specifically with a known immobilized ligand. In this study, by using the Biacore-X instrument, about 0.1 pmol β 2-microglobulin was immobilized on sensor chip, the interacting protein in solution was delivered to the biosensor chip surface. Then, a microrecovery method of the principle of “sandwich” structure was used to elute the bonding microglobulin protein from the sensor chip, and the protein identification was then achieved after tryptic digestion by matrix-assisted laser desorption/ionization-time of flight mass fingerprint mapping and data-base search. The strategy was successfully applied to the model protein microglobulin interacting with its antibody, a unambiguous identification was obtained at 10 -15 mol level.

关 键 词：生物传感芯片质谱 表面等离子共振 生物芯片 蛋白质组学 

分 类 号：Q657[生物学—生物物理学]