抗狂犬病毒单克隆抗体的研制及应用

GENERATION AND APPLICATION OF THE MONOCLONAL ANTIBODIES TO RABIES VIRUS

应用杂交瘤技术获得4株分泌抗狂犬病毒（Rabies virus,RV）单克隆抗体细胞株,并对其特性进行分析。经鉴定,它们所分泌的抗体类型为IgG1,腹水效价为1∶105~1∶107,ELISA法和间接免疫荧光检测（indirect immunofluoreseent assay,IFA）结果表明：该4株单抗对RV有良好的特异性,对正常Vero细胞及人白蛋白和其他细胞培养物无非特异性反应。将这些单抗腹水进行Protein G亲和层析纯化后获得的纯化抗体用于制备反相亲和层析柱,并以反相亲和层析法纯化RV以获得高纯度RV抗原。应用纯化的RV建立胶体金免疫层析法（gold-immunochromatography assay,GICA）检测RV抗体血清,结果显示,GICA与标准ELISA法检测比较的总符合率为96.1%。

Four hybridoma cell lines secreting monoclonal antibodies（McAbs） to rabies virus were established and the McAbs were characterized.Subclass of these McAbs was IgG1.The titers of ascitic fluids were 1×105-1×107.In indirect ELISA and IFA tests,all of four McAbs specifically reacted with rabies virus but did not react with Vero cells,human albumin and other cell cultures.The McAbs were purified by using protein G affinity chromatograph,which were then used to prepare anti-phase affinity chromatograph to purify rabies virus.The rabies virus preparation was used to develop gold-immunochromatography-assay（GICA） for detection of rabies antibodies in the serum samples.The comparative study showed 96.1% agreement between GICA and ELISA method.