【摘要】 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。更多还原

【Abstract】 Objective To construct human-mouse chimeric antibody against TNF-α and express in CHO cells.Methods Clone the gene encoding variable region of immunoglobulin from murine hybridoma cells by RT-PCR and RACE methods.Construct human-mouse chimeric antibody by fusing the cloned gene with the gene encoding the constant region of human immunoglobulin.Transfect CHO cells with the fusion gene and detect the expressed product.Results The primary expression level of the constructed chimeric antibody in CHO cells was 0.71 μg/ml,and that after subclone was 0.95 μg/ml.Conclusion Anti-human TNF-α chimeric antibody gene was successfully constructed and highly expressed in CHO cells.更多还原