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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.
In this paper, baculovirus expression system was used to express CEA murine-human chimeric antibody heavy and light chain genes in insect cells (Sf_9 and fall armyworm cells) The cloned heavy and light chain variable region (V_H, Vk) genes were respectively cloned with human immunoglobulin constant region genes Cr_3 and Ck to construct the chimeric antibody gene of chickens.The chimeric antibody gene transfer vector and linear The viral DNA was co-transfected into Sf_9 cells and the recombinant viruses were obtained by dot blot, PCR amplification and Southern blot analysis.Western blot analysis showed that chimeric heavy chain and light chain were expressed respectively in Sf_9 cells infected with recombinant virus, and radioimmunoassay The results of the RIA and indirect ELISA assays showed that chimeric heavy and light chain gene expression products have the ability to bind to CEA.