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针对脊髓灰质炎病毒非衣壳蛋白的单克隆和单特异性抗体的制备。

Production of monoclonal and monospecific antibodies against non-capsid proteins of poliovirus.

作者信息

Pasamontes L, Egger D, Bienz K

出版信息

J Gen Virol. 1986 Nov;67 ( Pt 11):2415-22. doi: 10.1099/0022-1317-67-11-2415.

Abstract

Non-capsid poliovirus proteins of the P2 region in extracts of infected cells were solubilized by SDS, separated by PAGE, electroeluted from the gels and used to immunize mice. The sera obtained were rendered monospecific by extensive absorption with proteins from uninfected cell extracts, and the spleen cells of these animals were used to establish antibody-secreting hybridomas. The monoclonal and the monospecific antibodies recognized denatured antigen (proteins 2C, 2BC and P2) in ELISA, immunoblot and immunoprecipitation tests and also combined with the native proteins of the P2 region in immunoprecipitation. In addition, the antibodies could be used successfully for immunofluorescence and electron microscopic immunocytochemistry.

摘要

感染细胞提取物中P2区域的非衣壳脊髓灰质炎病毒蛋白经SDS溶解,通过聚丙烯酰胺凝胶电泳(PAGE)分离,从凝胶中电洗脱出来并用于免疫小鼠。通过用未感染细胞提取物中的蛋白质进行广泛吸收,使获得的血清具有单特异性,这些动物的脾细胞用于建立分泌抗体的杂交瘤。单克隆抗体和单特异性抗体在酶联免疫吸附测定(ELISA)、免疫印迹和免疫沉淀试验中识别变性抗原(蛋白2C、2BC和P2),并且在免疫沉淀中也与P2区域的天然蛋白结合。此外,这些抗体可成功用于免疫荧光和电子显微镜免疫细胞化学。

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