The 38 kDa protein is a major antigen of mycobacterium tuberculosis and has been widely used in TB serodiagnosis, due to its highly sensitivity and specificity. Here we attempt to establish a production platform of recombinant 38 kDa protein in mammalian cells and to evaluate the potential value of 38 kDa protein in TB serodiagnosis. The 38 kDa gene is synthesized and cloned into a lentiviral expressing vector. Recombinant lentiviral vector LV-CMV-38 kDa-eGFP was packaged, titered, and then transduced into HEK 293 T cells. Recombinant cell lines were selected by limiting dilution. Supernatants were collected and purified by HisTrapTM HP column. Western blot showed a molecular weight of approximate 38 kDa in cell supernatants as expected. ELISA assay confirmed the immunological specificity of the obtained protein in the presence of MTB-infected human serum samples. In all, we have obtained a stable cell line with long-term and robust expression of secretory MTB 38 kDa protein, which may provide a promising candidate antigen for the development of TB serological diagnosis.

38 kDa 蛋白是结核分枝杆菌的主要抗原，由于其高度敏感性和特异性，已广泛用于结核病血清学诊断。在这里，我们试图在哺乳动物细胞中建立重组 38 kDa 蛋白的生产平台，并评估 38 kDa 蛋白在结核病血清学诊断中的潜在价值。合成 38 kDa 基因并克隆到慢病毒表达载体中。将重组慢病毒载体 LV-CMV-38 kDa-eGFP 包装、滴定，然后转导到 HEK 293 T 细胞中。通过有限稀释选择重组细胞系。上清液通过 HisTrapTM HP 柱收集和纯化。正如预期的那样，蛋白质印迹显示细胞上清液中的分子量约为 38 kDa。ELISA 测定证实了在 MTB 感染的人血清样品存在下获得的蛋白质的免疫特异性。总之，我们获得了一个稳定的细胞系，该细胞系长期稳定表达分泌型 MTB 38 kDa 蛋白，可能为结核病血清学诊断的发展提供有希望的候选抗原。