CAJ | 학술논문

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。
목적：제비유항인류표피생장인자수체2（HER-2／NEU）단극륭항체곡타주단항（Trastuzumab，간칭 T）여남과단백（Cucurmosin，간칭CUS）우련이성적면역독소，병진행감정。방법이N-호박선알-3-（2-필정이류）병산지（ SPDP）위우련제，제비 T-CUS，경과Ni Sepharose 6 Fast Flow화SP Sepharose 6 Fast Flow순화후SDS-PAGE분석기순도급성분，세포ELISA법검측기대인유선암BT-474세포적결합능력。결과 SDS-PAGE분석현시T-CUS면역독소성공구건，면역독소기본보류료대BT-474세포적결합능력。결론이용SPDP가실현곡타주단항여CUS적성공우련。
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.