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目的构建抗人TNF-α人-鼠嵌合抗体基因,并在CHO细胞中表达。方法采用RT-PCR及RACE技术从鼠杂交瘤细胞克隆免疫球蛋白可变区基因,与人免疫球蛋白恒区基因拼接,构建人-鼠嵌合抗体基因,转染CHO细胞并测定其表达量。结果构建的嵌合抗体基因在CHO细胞中的初始表达量为0.71μg/ml,亚克隆后表达量达0.95μg/ml。结论已成功构建抗人TNF-α嵌合抗体基因,并在CHO细胞中得到了高效表达。
Objective To construct an anti-human TNF-α human-mouse chimeric antibody gene and express it in CHO cells. Methods The immunoglobulin variable region genes were cloned from murine hybridoma cells by RT-PCR and RACE techniques. The human immunoglobulin constant region gene was spliced to construct the human-mouse chimeric antibody gene. The recombinant plasmid was transfected into CHO cells and its expression level . Results The initial expression level of chimeric antibody gene was 0.71μg / ml in CHO cells and 0.95μg / ml in subclone. Conclusion The chimeric antibody against human TNF-α has been successfully constructed and has been highly expressed in CHO cells.