抗禽流感病毒H5N1亚型单克隆抗体制备初报

目的制备禽流感H5N1亚型病毒的单克隆抗体,为相关研究提供工具。方法以禽流感H5N1亚型病毒免疫BALBc小鼠,取其脾细胞和SP20细胞融合,用血凝抑制试验(HI)和酶联免疫反应(ELISA)检测培养上清,并将阳性融合细胞稀释克隆化3次直至100%孔均为阳性,筛选阳性克隆株,运用免疫荧光法评估单克隆抗体检测病毒感染的犬肾细胞(MDCK)。结果得到三株稳定分泌抗体的细胞并命名为F8、F9、G11,抗体亚型鉴定结果分别为IgG1、IgG2a和IgG2b;在免疫荧光法单克隆抗体能够检测出感染MDCK细胞的病毒。结论建立了3株抗禽流感H5N1亚型病毒的单克隆抗体细胞株,其产生的一株高特异性的McAbG11能够用于H5N1亚型禽流感病毒感染诊断,并可能应用于禽流感病毒H5N1亚型感染的防治。

Preparation of Monoclonal Antibodies Specific to Subtype H5N1 of Avian Influenza Virus

Objective To prepare the monoclonal antibodies specific to subtype of H5N1 avian influenza virus and offer tool for correlative researches.Methods Monoclonal antibodies specific to subtype of H5N1 avian influenza virus were developed by fusing SP2/0 myeloma cell with spleen cell of BALB/c mice immunized with AIV_H5N1, the characterization of three McAbs can specifically react with AIV_H5N1 by hemagglutination inhibit test(HI), enzyme linked immunosorbent assay(ELISA) and immunofluorescence. The inmmunoglobulin subclass of the McAbs was then identified. Results Three colone hybridoma secreting McAbs against AIV_H5N1 were obstained as F8,F9,G11 respectively, the inmmunoglobulin subclass of the McAbs was identified as IgG1,IgG2a and IgG2b. Conclusion The highly effective and specific McAbs G11 can play an important role in diagnosis, treatment and prevention of the infection of AIV_H5N1