免疫毒素Trastuzumab-Cucurmosin（T-CUS）的制备及鉴定

摘要

OBJECTIVE To prepare an immunotoxin ( IT) by coupling the monoclonal antibody against human epidermal growth factor receptor ( HER-2/NEU) to Cucurmosin( CUS) ,then identify.METHODS T-CUS was pre-pared using N-succinimidyl-3-(2-pyridyldithio)-propionate ( SPDP) as a coupler,and was purified using Ni Sepha-rose 6 Fast Flow column and SP Sepharose 6 Fast Flow column, then analyzed for purity and component by SDS-PAGE,and determined for binding ability to human breast cancer BT-474 cells by ELISA.RESULTS SDS-PAGE showed that T-CUS was successfully constructed,and the binding ability of human breast cancer BT-474 cells was ba-sically retained.CONCLUSION Trastuzumab may be successfully coupled to CUS by using SPDP.%目的：制备由抗人类表皮生长因子受体2（HER-2／NEU）单克隆抗体曲妥珠单抗（Trastuzumab，简称 T）与南瓜蛋白（Cucurmosin，简称CUS）偶联而成的免疫毒素，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 T-CUS，经过Ni Sepharose 6 Fast Flow和SP Sepharose 6 Fast Flow纯化后SDS-PAGE分析其纯度及成分，细胞ELISA法检测其对人乳腺癌BT-474细胞的结合能力。结果 SDS-PAGE分析显示T-CUS免疫毒素成功构建，免疫毒素基本保留了对BT-474细胞的结合能力。结论利用SPDP可实现曲妥珠单抗与CUS的成功偶联。