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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 and identify immunotoxins (IT) derived from the conjugation of monoclonal antibody (McAb) against epidermal growth factor receptor (EGFR) with ricin A chain (RTA). Methods The immunotoxin EGFRmab-RTA was prepared by using N-succinamide-3 (2-pyridyldithio) propionate (SPDP) as coupling agent. The purity and composition of the immunotoxin were determined by SDS-PAGE. Human hepatoma HepG2 cell binding ability. Results SDS-PAGE analysis showed that the immunotoxin was successfully constructed. The conjugation ratio of EGFRmab to RTA was 1: 1.16. Immunotoxin basically retained the binding ability of EGFRmab to HepG2 cells. Conclusion The successful coupling of EGFRmab with RTA can be achieved by using SPDP.