Li R, Xue A, Zhu X, Cai S
Biotechnology Research Center, CAAS (Chinese Academy of Agricultural Sciences), Beijing.
Chin J Biotechnol. 1992;8(4):277-81.
Two hybridomas stably secreting monoclonal antibodies (McAb) against sweet potato feathery mottle virus (SPFMV) were constructed by fusing mouse myeloma (Sp2/0-Ag14) with mouse splenocytes immunized with SPFMV-LF. The McAb secreted by the two hybridomas specifically reacted with both SPFMV-LF (China strain) and SPFMV-RC (American strain) in dot-blot-ELISA. A large amount of McAb was produced with BALB/c mice. The titer of hybridoma cultured supernatant was 1:256, and that of the ascitic fluids was 1:1 x 10(4); and the immunoglobulins of McAb were IgG2a and IgG3 respectively. The detection of SPFMV-infected sweet potato plants with McAb confirmed that the McAb obtained in this test were desirable for large-scale routine surveys of SPFMV. This study on construction of hybridomas and detection of SPFMV using McAb is the first report in the world.
通过将小鼠骨髓瘤细胞(Sp2/0-Ag14)与经甘薯羽状斑驳病毒-LF(SPFMV-LF)免疫的小鼠脾细胞融合,构建了两株稳定分泌抗甘薯羽状斑驳病毒(SPFMV)单克隆抗体(McAb)的杂交瘤细胞。在斑点印迹ELISA中,这两株杂交瘤细胞分泌的单克隆抗体与SPFMV-LF(中国株)和SPFMV-RC(美国株)均发生特异性反应。用BALB/c小鼠制备了大量单克隆抗体。杂交瘤细胞培养上清液的效价为1:256,腹水的效价为1:1×10⁴;单克隆抗体的免疫球蛋白分别为IgG2a和IgG3。用单克隆抗体检测感染SPFMV的甘薯植株,证实本试验获得的单克隆抗体适用于SPFMV的大规模常规检测。关于杂交瘤细胞构建及使用单克隆抗体检测SPFMV的这项研究是世界上的首次报道。