蓖麻蛋白A链与单抗3G11偶联物的制备及体外生物活性研究  

The preparation of immunoconjugate composed of ricin A-chain and mAB 3G11 and the evaluation of its biological activity in vitro

作　　者：宋波[1] 戴垚[2] 黎维勇[1] 金文闻[3] 陈华庭[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,湖北武汉430022 [2]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050 [3]华中科技大学生命学院资源生物学与生物技术研究室,湖北武汉430014

出　　处：《中国医院药学杂志》2005年第10期899-901,共3页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金资助项目(编号:30271552)

摘　　要：目的:Ⅳ型胶原酶在肿瘤生长和转移的多个环节发挥着重要作用。将抗Ⅳ型胶原酶单克隆抗体3G11与蓖麻毒素A链(RA)偶联形成免疫毒素(ITs),并评价了该偶联物的体外生物活性。方法:从蓖麻籽中提取纯RA,与N-琥珀酰亚胺基3-(2-吡啶二硫基)丙酸酯(SPDP)修饰的单抗3G11偶联,进一步纯化后得到3G11-RA。以ELISA实验测定ITs的免疫反应性,并通过四甲基偶氮唑盐(MTT)实验对其体外细胞毒性进行评价。结果:ITs基本保持了单抗3G11对人肝癌细胞BEL-7402的免疫反应性,其对BEL-7402的细胞毒性比RA显著增强近50倍,而对人正常肝细胞LO2的毒性与RA相似。结论:3G11-RA免疫毒素对BEL-7402肝癌细胞具有较好的免疫反应性和体外细胞毒性。OBJECTIVE Type Ⅳ collagenase plays an important role in many steps of tumor growth and metastasis. The aim of this study was to evaluate the biological activity of the 3G11-RA immunoconjugate in vitro composed by ricin A-chain and anti-type IV collagenase monoclonal antibody （mAb） 3G11. METHODS mAb 3G11 modified by SPDP was conjugated with ricin A-chain, which was purified from castor beans. After further purification, the immunoreactivity and cytotoxicity of 3G11-RA was determined by ELISA and MTT method, respectively. RFSULTS The immunoreactivity of ITs with BEL-7402 tumor cells is almost the same as free 3G11. The cytotoxIcity of ITs to BEL-7402 is nearly 50-times that of ricin A-chain. But the ITs＇ cytotoxicity to LO2 normal liver cell is similar as ricin A-chain. CONCLUSION The ITs showed high immunoreactivity and cytotoxicity to the BEL-7402 tumor cells in vitro.

关 键 词：单克隆抗体 蓖麻毒素A链 免疫毒素 体外细胞毒性 肿瘤生长 

分 类 号：R197.322[医药卫生—卫生事业管理] R288[医药卫生—公共卫生与预防医学]