表面等离子共振测定CD45-FITC抗体与蛋白A特异性结合常数  

Determination of the specific binding constants of CD45-FITC to protein A by SPR

作　　者：张晖[1] 薛洪宝[1] 陈飞剑[1] 王杰[1] 王充[2] 李柏青[2] ZHANG Hui;XUE Hongbao;CHEN Feijian;WANG Jie;WANG Chong;LI Baiqing(Department of Chemistry,Bengbu Medical College,Bengbu 233030,China;Anhui Key Laboratory of Infection and Immunity∥Bengbu Medical College,Bengbu 233030,China)

机构地区：[1]蚌埠医学院化学教研室,安徽蚌埠233030 [2]感染与免疫安徽省重点实验室∥蚌埠医学院,安徽蚌埠233030

出　　处：《中山大学学报（自然科学版）》2018年第4期145-149,共5页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金青年项目(21601004);安徽高校自然科学研究重点项目(KJ2016A485);安徽高校科研平台创新团队建设项目(2016-40)

摘　　要：运用表面等离子共振(SPR)平台建立测定CD45-FITC抗体与蛋白A特异性结合常数的方法。当偶联蛋白A的传感芯片上流过CD45-FITC抗体溶液时,双通道SPR仪实时监测CD45-FITC抗体与蛋白A特异性结合过程,获得了该反应不同温度下的结合速率常数k_a、解离速率常数k_d和结合平衡常数K_a,以及该反应的活化能E_a和焓变△H。结果表明该特异性结合比较适宜的条件是:中性或微碱性、较高的离子强度和较高的温度。该方法操作步骤简单、快速、灵敏度高、消耗样品少,是研究分析免疫球蛋白IgG抗体和蛋白A等生物分子相互作用比较理想的方法之一。A SPR method was established to determine the specific binding constants of CD45-FITC to protein A. The specific binding of CD45-FITC was monitored in real time on the SR7500DC Dual Channel System. The characteristic constants for this reaction they were association rate constant k a, dissociation rate constant k d, association equilibrium constant K a,activation energy E a and enthalpy change △ H . The results indicated that the optimum reaction conditions were under neutral or alkaline medium, with a slightly higher ionic strength and temperature. This method has the characteristics of being fast, simple operation, high sensitivity and less sample, which can be suitable for the investigation of biomolecular interactions of immunoglobulin IgG with protein A.

关 键 词：表面等离子共振 CD45-FITC抗体 蛋白A 特异性结合常数 

分 类 号：O657.39[理学—分析化学]