摘要
目的制备蓖麻毒蛋白（Ricin）A链（RTA）与转铁蛋白受体（TFR/CD71）mAb偶联的免疫毒素（IT），并考察其体外靶向抗肿瘤活性。方法以蓖麻子为原料，提纯RTA，在偶联剂N-琥珀酰胺-3-（2-吡啶二硫）丙酸酯（SPDP）的作用下，构建IT；并以间接ELISA法检测IT的免疫反应性，以CCK-8法测定并比较IT在体外对人肝癌SMMC-7721细胞和正常肝细胞LO2的细胞毒效应。结果SPDP偶联法制备的IT对肿瘤细胞的免疫反应性无明显变化，体外实验结果显示，各浓度IT对SMMC-7721细胞的杀伤作用较LO2细胞差异具有统计学意义（P＜0.05）。结论CD71mAb-RTA免疫反应性稳定，对肿瘤和正常体细胞的体外细胞毒效应差异明显，提示其对肿瘤细胞有较好的靶向杀伤作用，有望开发成为肿瘤的靶向治疗药物。
Objective To prepare the immune toxin(IT)of Ricin A chain(RTA)and transferrin receptor(TFR/CD71)mAb coupling,and to investigate the anti-tumor activity in vitro.Methods IT was constructed by using castor beans as raw materials,purified RTA,under the action of the coupling agent N-succinamide-3-(2-pyridine disulfide)propionate(SPDP).In order to detect the immune reactivity of IT in the indirect ELISA method,the cck-8method was used to determine and compare the cytotoxic effects of IT in vitro for human liver cancer cell SMMC-7721and normal hepatocellular LO2.Results SPDP coupling method of IT there was no significant alteration of the immune response to tumor cells,in vitro experimental results showed that the concentration IT on LO2cells SMMC-7721cell killing effect significantly enhanced(P<0.05).Conclusion CD71mAb-RTA immune reactivity is stable,and there was obvious difference betweem the tumor and normal cells in vitro cytotoxic effect.It has better targeted killing effect on tumor cells,and is expected to develop into tumor targeted therapy drugs.
作者
陈海霞
陈祥胜
黄婧
祝香芝
CHEN Hai-xia;CHEN Xiang-sheng;HUANG Jing;ZHU Xiang-zhi(Hubei University of Traditional Chinese Medicine, Wuhan 430065, China)
出处
《中国生化药物杂志》
CAS
2017年第11期1-3,6,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
湖北省卫计委资助项目(WJ2015MB132)
湖北省教育厅科研项目(B2013129)
