免疫毒素EGFR mab-RTA的制备及鉴定  

Preparation and Identification of Immunotoxin EGFR mab-RTA

作　　者：申苏建[1] 吴金明[1] 金思思[1] 黄智铭[1] 吴建胜[1] 

机构地区：[1]温州医学院附属第一医院消化内科,浙江温州325000

出　　处：《中国生物制品学杂志》2010年第5期510-513,共4页Chinese Journal of Biologicals

摘　　要：目的制备由抗表皮生长因子受体(EGFR)单克隆抗体(mab)与蓖麻毒蛋白A链(RTA)偶联而成的免疫毒素(IT),并进行鉴定。方法以N-琥珀酰胺-3(-2-吡啶二硫)丙酸酯(SPDP)为偶联剂,制备免疫毒素EGFRmab-RTA,SDS-PAGE分析其纯度及成分,DAB显色法检测其对人肝癌HepG2细胞的结合能力。结果 SDS-PAGE分析显示免疫毒素成功构建,偶联物中EGFRmab与RTA的偶联比为1:1.16,免疫毒素基本保留了EGFRmab对HepG2细胞的结合能力。结论利用SPDP可实现EGFRmab与RTA的成功偶联。Objective To prepare an immunotoxin（IT）by coupling the monoclonal antibody（mab）against epidermal growth factor receptor（EGFR）to ricin A（RTA）,then identify.Methods IT EGFR mab-RTA was prepared using N-succinimidyl-3（-2pyridyldithio）-propionate（SPDP）as a coupler,then analyzed for purity and component by SDS-PAGE,and determined for binding ability to human hepatocarcinoma HepG2 cells by DAB coloration.Results SDS-PAGE showed that IT EGFR mab-RTA was successfully constructed,in which the coupling ratio of EGFR mab to RTA was 1：1.16,and the binding ability of EGFR mab to HepG2 cells was basically retained.Conclusion EGFR mab may be successfully coupled to RTA by using SPDP.

关 键 词：表皮生长因子受体 单克隆抗体 蓖麻毒蛋白A链 免疫毒素 

分 类 号：R392.33[医药卫生—免疫学]