Qiu Zheng, Tian Wei, Yu Tianfei, Li Li, Ma Bo, Wang Junwei
College of Veterinary Medicine, Northeast Agricultural University, Xiangfang District, Harbin, China.
Hybridoma (Larchmt). 2012 Apr;31(2):125-30. doi: 10.1089/hyb.2011.0098.
In the present study, monoclonal antibodies (MAbs) against NS1 protein of Goose parvovirus (GPV) were generated. The secreted MAbs were obtained by fusing mouse myeloma cells and spleen cells of BALB/c mice, which were immunized with the plasmid pcDNA3.1-GPV-NS1 and recombinant protein of GPV-NS1. With indirect ELISA, six hybridoma cell lines against GPV-NS1 were screened. The subtypes of the two MAbs were IgG2a; the others were IgM. The light chain was κ. Western blot analysis showed that six MAbs reacted with recombinant protein GPV-NS1. GPV-NS1 was dissected into 15 overlapping epitopes, which were used to react with MAbs in Western blot. Results showed that six MAbs recognized NS1 protein linear B-cell epitopes located at the C-terminus 453-514 aa, 485-542 aa, and 533-598 aa.
在本研究中，制备了针对鹅细小病毒（GPV）NS1蛋白的单克隆抗体（MAb）。通过将小鼠骨髓瘤细胞与用质粒pcDNA3.1-GPV-NS1和GPV-NS1重组蛋白免疫的BALB/c小鼠的脾细胞融合，获得分泌型单克隆抗体。采用间接ELISA法筛选出6株抗GPV-NS1的杂交瘤细胞系。其中两株单克隆抗体的亚型为IgG2a；其他为IgM。轻链为κ链。Western blot分析表明，6株单克隆抗体与重组蛋白GPV-NS1发生反应。将GPV-NS1切割成15个重叠表位，用于在Western blot中与单克隆抗体反应。结果表明，6株单克隆抗体识别位于C末端453-514 aa、485-542 aa和533-598 aa的NS1蛋白线性B细胞表位。
