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为制备狂犬病病毒糖蛋白(G蛋白)特异性单克隆抗体(McAb)及鉴定其识别范围,采用人工合成多肽“PPDQLVNLHDFRSDEIEHLVVEE”与KLH的偶联物免疫小鼠,经筛选制备出1株阳性杂交瘤细胞,记作4D3,其亚类鉴定为IgG2b/κ链。构建了真核表达载体pCAGGS-G/pCAGGS-13G,经鉴定可正确表达G蛋白,用于鉴定McAb。免疫荧光与Western-blot分析显示,该株McAb与真核表达的人工减毒株LBNSE G蛋白可发生特异反应,而与强毒株IMDRV-13的G蛋白不反应。本试验结果可为进一步研究G蛋白的结构、功能和毒株的初步筛查提供参考。
In order to prepare specific monoclonal antibody (McAb) of rabies virus glycoprotein (G protein) and identify its identification range, a mouse immunized with the conjugate of synthetic peptide “PPDQLVNLHDFRSDEIEHLVVEE ” and KLH was used to prepare a positive Hybridoma cells, designated 4D3, whose subclasses were identified as IgG2b / kappa chain. The eukaryotic expression vector pCAGGS-G / pCAGGS-13G was constructed and correctly identified to express G protein for McAb identification. Immunofluorescence and Western-blot analysis showed that the McAb specifically reacted with the eukaryotic expressed artificial attenuated strain LBNSE G protein and did not react with the G protein of the virulent strain IMDRV-13. The results of this study can provide a reference for further study on the structure, function and preliminary screening of G protein.