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分泌抗甘薯羽状斑驳病毒单克隆抗体的杂交瘤细胞系的构建及其抗体在甘薯羽状斑驳病毒检测中的应用

Construction of hybridomas secreting monoclonal antibody against sweet potato feathery mottle virus and use of antibody for detection of SPFMV.

作者信息

Li R, Xue A, Zhu X, Cai S

机构信息

Biotechnology Research Center, CAAS (Chinese Academy of Agricultural Sciences), Beijing.

出版信息

Chin J Biotechnol. 1992;8(4):277-81.

PMID:1343832
Abstract

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.

摘要

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。

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