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用宫颈癌细胞Hela表面高表达G250抗原的单克隆抗体G250修饰非病毒基因载体,获得肿瘤靶向基因载体.通过注射G250杂交瘤细胞于小鼠腹腔,制备富含G250mAb的腹水,用正辛酸-硫酸铵沉淀法和Protein A Agarose分离纯化,获得高纯度的G250mAb.通过二硫键将PEI与G250mAb偶联,得到修饰的基因载体G250mAb-PEI,研究其转基因靶向性.结果表明,G250mAb-PEI对Hela细胞的基因转染具有显著的靶向性,对Hela细胞的转基因效率是肝癌细胞HepG2(G250阴性)的2倍;而对正常血管平滑肌细胞(SMC)的基因转染效率比Hela低近20倍,G250mAb修饰与否对SMC没有靶向性;对3T3细胞的毒性显著低于未修饰的PEI,表明G250mAb-PEI是一种高效、低毒和具有靶向性的基因载体.
The non-viral gene vector was modified with monoclonal antibody G250 highly expressing G250 antigen on the Hela surface of cervical cancer cells to obtain the tumor-targeted gene vector.The G250mAb-ascitic fluid was prepared by injecting G250 hybridoma into abdominal cavity of mice, Ammonium sulfate precipitation and Protein A Agarose to obtain high purity G250mAb.The PEI was coupled with G250mAb by disulfide bond to obtain the modified gene carrier G250mAb-PEI, and its transgene targeting was studied.The results showed that G250mAb-PEI Hela cell gene transfection has a significant target of Hela cells transgene efficiency HepG2 (G250 negative) 2 times; and normal vascular smooth muscle cells (SMC) gene transfection efficiency lower than Hela 20-fold, G250mAb modification SMC no targeting; 3T3 cells was significantly lower toxicity than unmodified PEI, indicating that G250mAb-PEI is a highly efficient, low-toxic and targeted gene vector.