Ebihara Takeru, Xu Jian, Tonooka Yoshino, Nagasato Takumi, Kakino Kohei, Masuda Akitsu, Minamihata Kosuke, Kamiya Noriho, Nakatake Hirokazu, Chieda Yuuka, Mon Hiroaki, Fujii Tsuguru, Kusakabe Takahiro, Lee Jae Man
Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, Fukuoka 819-0395, Japan.
Laboratory of Biology and Information Science, Biomedical Synthetic Biology Research Center, School of Life Sciences, East China Normal University, Shanghai 200062, China.
Insects. 2021 Jun 2;12(6):517. doi: 10.3390/insects12060517.
The tumor necrosis factor α (TNFα) has been employed as a promising reagent in treating autoimmunity and cancer diseases. To meet the substantial requirement of TNFα proteins, we report in this study that mature types of recombinant human and murine TNFα proteins are successfully expressed in the baculovirus expression system using silkworm larvae as hosts. The biological activities of purified products were verified in culture murine L929 cells, showing better performance over a commercial -derived murine TNFα. By comparing the activity of purified TNFα with or without the tag removal, it is also concluded that the overall activity of purified TNFα cytokines could be further improved by the complete removal of C-terminal fusion tags. Collectively, our current attempt demonstrates an alternative platform for supplying high-quality TNFα products with excellent activities for further pharmaceutical and clinical trials.
肿瘤坏死因子α（TNFα）已被用作治疗自身免疫性疾病和癌症的一种有前景的试剂。为满足对TNFα蛋白的大量需求，我们在本研究中报告，以家蚕幼虫为宿主，在杆状病毒表达系统中成功表达了成熟型重组人源和鼠源TNFα蛋白。在培养的小鼠L929细胞中验证了纯化产物的生物活性，结果表明其性能优于市售鼠源TNFα。通过比较去除标签和未去除标签的纯化TNFα的活性，还得出结论，完全去除C端融合标签可进一步提高纯化TNFα细胞因子的整体活性。总体而言，我们目前的尝试展示了一个替代平台，可为进一步的药物和临床试验提供具有优异活性的高质量TNFα产品。
