Meng Fandan, Yin Jiechao, Li Xunliang, Yang Wei, Li Guangxing, Ren Xiaofeng
College of Veterinary Medicine, Northeast Agricultural University, Harbin, China.
Hybridoma (Larchmt). 2010 Aug;29(4):345-50. doi: 10.1089/hyb.2010.0009.
Transmissible gastroenteritis virus (TGEV) is a member of coronaviruses. The viral spike (S) protein mediates the interaction between TGEV and its susceptible cells. Here, we expressed a truncated gene encoding the N terminal half of TGEV S gene (designated S1 gene) in a prokaryotic system. The resulting S1 protein was used to immunize BALB/c mice followed by the generation of a monoclonal antibody (MAb). A generated MAb (7F9) was identified by ELISA and the chromosome number of the hybridoma cell was analyzed. The immunoreactivity of the MAb to TGEV S protein was confirmed by Western blot analysis. Moreover, immunofluorescence assays showed that the MAb is able to detect cell infection by TGEV. The MAb achieved in this study can be used as a specific diagnostic reagent for detecting TGEV S protein.
传染性胃肠炎病毒（TGEV）是冠状病毒的一种。病毒刺突（S）蛋白介导TGEV与其易感细胞之间的相互作用。在此，我们在原核系统中表达了一个编码TGEV S基因N端一半的截短基因（命名为S1基因）。所得的S1蛋白用于免疫BALB/c小鼠，随后产生单克隆抗体（MAb）。通过ELISA鉴定产生的单克隆抗体（7F9），并分析杂交瘤细胞的染色体数目。通过蛋白质印迹分析证实了该单克隆抗体对TGEV S蛋白的免疫反应性。此外，免疫荧光试验表明该单克隆抗体能够检测TGEV引起的细胞感染。本研究中获得的单克隆抗体可作为检测TGEV S蛋白的特异性诊断试剂。
