H5亚型禽流感病毒血凝素蛋白单克隆抗体的制备及鉴定

Preparation and Identification of Monoclonal Antibodies Against Haemagglutinin of Subtype H5 Avian Influenza Virus

用纯化的H5亚型禽流感病毒(AIV)抗原免疫Balb/C小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞融合,经筛选,获得9株稳定分泌单克隆抗体的杂交瘤细胞株,其中4株能高效分泌血凝素(HA)蛋白特异性的单克隆抗体.特异性试验表明,IE6单克隆抗体仅与试验的H5病毒株反应,而不与新城疫病毒(NDV)、H9亚型AIV和产蛋下降综合症病毒(EDSV)反应.这9株单克隆抗体分别为IgG1、IgG2a、IgG2b、IgM亚类,κ轻链.

Balb/C mice were immunized with the purified H5N1 and H9N2 subtype avian influenza virus（AIV） antigens.And monoclonal antibodies against above antigens were produced.Splenocytes from the immunized mice were fused with SP2/0 myeloma cells.Positive hybridoma clones were screened by ELISA,and nine monoclonal antibodies were generated following conventional protocols for making monoclonal antibodies.The nine monoclonal antibodies reacted with newcastle disease virus（NDV）,H5 subtype and H9 subtype AIV antigen,and only IE6 was specific to H5 subtype AIV.And IIB6C3,IIA8H11,IE6,IIIC2D1 were specific to the hemagglutinin（HA） fusion proteins.The nine monoclonal antibodies belonged to the IgG1、IgG2a、IgG2b、IgM subclass,with κ type light chain.