抗癌胚抗原鼠人嵌合抗体重、轻链基因在昆虫细胞中的表达

Separate Expression of A Murine-Human Chimeric Antibody Heavy and Light Chain Genes with Specificity for Carcinoembryonic Antigen in Insect Cells

本文采用杆状病毒表达系统在昆虫细胞(Sf_9,秋粘虫细胞)中表达了抗癌胚抗原(CEA)鼠-人嵌合抗体重、轻链基因.将鼠源性单抗杂交瘤细胞中已克隆到的重、轻链可变区(V_H、Vk)基因分别与人免疫球蛋白恒定区基因Cr_3、Ck相拼接,构建鼠人嵌合抗体基因.含嵌合抗体基因的转移载体与线性化病毒DNA共转染Sf_9细胞,并通过点杂交、PCR扩增和Southern blot分析获得重组病毒.Western blot分析表明以重组病毒感染的Sf_9细胞分别表达了嵌合重链和轻链,放射免疫分析法(RIA)和间接ELISA测定的结果表明嵌合重、轻链基因表达产物具有与CEA结合的能力.

The heavy and light chain genes of a murine-human chimeric antibody with specificity for carcinoembryonic antigen have been expressed separately in a baculovirus expression system.The genes encoding the heavy and light chain variable region (VH and Vk) were combined to the human Cr3 and Ck genes respectively to construct the murine-human chimeric antibody genes. The transfer vectors containing the chimeric genes were contransferted into Sf9 cells with linerized virus DNA and recombinant virus were isolated by dot hybridization PCR and southern blot analysis. Western blot showed the chimeric heavy and light chain were expressed in the recombinant virus infected Sf9 cells. RIA and ELISA demonstrated that both of the expressed chimeric heavy and light chain have the CEA-binding abilities.