可溶性重组蛋白EGF-Ang构建及体外细胞毒性检测  

Construction and Cytotoxicity In Vitro of Soluble Fusion Protein EGF-Ang

作　　者：岳玉环[1] 朱平[1] 李丽萍[1] 吴广谋[1] 李铁征 何畅[1] 

机构地区：[1]解放军军需大学研究所,长春130062 [2]长春亚泰集团生物制药有限公司,长春130018

出　　处：《中国生物制品学杂志》2003年第1期16-18,共3页Chinese Journal of Biologicals

摘　　要：目的　构建人源化的免疫融合蛋白 ,以期获得具有较低免疫原性的靶向治疗药物。方法　利用基因工程技术将人表皮生长因子 (EGF)和人血管生成素 (Ang)基因连接起来 ,克隆到高效表达载体pET2 0b(+)中 ,构建重组表达质粒pET2 0 EA ,并在大肠杆菌中表达该融合蛋白 (EGF Ang)。用MTT法检测可溶性蛋白的细胞毒性作用。结果　经SDS PAGE和薄层扫描分析表明 ,外源蛋白表达量占菌体裂解蛋白总量的 6 .4 %。细胞活性检测表明EGF Ang重组蛋白在体外对过度表达EGFR的Hep2 和SP2 / 0细胞具有明显杀伤作用。结论　人源化免疫融合蛋白EGFObjective To construct an immune fusion protein from human origin and obtain a target anticarcinogen with low immunogenicity.Methods Epidermal growth factor(EGF) and angiogenin(Ang) genes were bound by genetic engineering technique and cloned into high expression vector pET20b(+) to construct a recombinant plasmid pT20 EA,then transformed to E.coli for expression.The cytotoxicity of expressed product was detected by MTT method.Results SDS PAGE and thin layer scanning showed that the expressed product contained about 6.4% of total somatic protein.Cytotoxicity ananlysis proved that the recombinant EGF Ang protein showed significant killing effect on the Hep2 and SP 2/0 cells overexpressing EGFR.Conclusion The construction of fusion protein EGF Ang laid a foundation of developing a target anticarcinogen.

关 键 词：表皮生长因子 血管生成素 可溶性重组蛋白EGF-Ang 体外细胞毒性 检测 

分 类 号：Q789[生物学—分子生物学] R730.5[医药卫生—肿瘤]