摘要
为了降低抗人胃癌鼠单抗3H11的免疫原性,以利于该单抗在临床中的应用。我们构建并表达了3H11的人-鼠嵌合抗体,将3H11的轻、重链可变区基因分别插入到含有人k链及IgGl重链恒定区基因的真核细胞表达载体中,构建了3H11人-鼠嵌合抗体轻、重链表达载体。应用Lipofectin方法先将嵌合轻链表达载体转染到Sp2/0细胞中,经用含霉酚酸的选择培养基筛选及克隆化培养,获得稳定分泌3H11人-鼠嵌合轻链的转染细胞株。再将嵌合重链表达载体转染该细胞系,用含有组氨醇的选择培养基筛选,获得组氨醇抗性细胞株,经亚克隆后得到可稳定分泌人k链和人IgGl的转染细胞系,经ELISA检测该细胞系所分泌的上清含有可与人胃癌细胞系803结合的人IgG抗体活性,RT-PCR结果显示该细胞株有人-鼠嵌合抗体mRNA的转录,证明已获得分泌3H11人-鼠嵌合抗体的细胞系。
In order to reduce its immunogenicity and increase its therapeutic usefullness, the murine anti-gastric cancer McAb 3H11 was genetically engineered into a human-mouse chimeric antibody. The 3H11 light and heavy chain variable genes were inserted into chimeric antibody expression vectors respectively. The chimeric light chain expression vector was first transfected into murine Sp2/0 myeloma cells using lipofectin. Transfectants that stably expressed human-mouse chimeric light chain were isolated by mycophenolic acid selection. Then the chimeric heavy chain expression vector was transfected into the transfectomas. Histidinol resistent cells were obtained and among these, clones stably secreting human-mouse chimeric antibodies were selected. The expression of human-mouse chimeric light and heavy chain mRNAs were proved by RT-PCR and the expressed chimeric antibody was demonstrated capable of binding to human gastric cancer cell MGC803
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1998年第2期116-120,共5页
Chinese Journal of Cancer Biotherapy
基金
北京市肿瘤分子生物学实验室项目
关键词
3H11
人-鼠嵌合抗体
胃肿瘤
真核基因
表达
human-mouse chimeric antibody
3H11 murine McAb
human gastric cancer
eukaryodc expression
