H5N1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 HA protein of AIV H5N1 subtype

为制备抗H5N1禽流感病毒(AIV)的单克隆抗体(MAb),本研究以灭活的H5N1亚型A/Chicken/Guangdong/S2261/2009(H5N1GD261)株免疫4周龄～6周龄BALB/c雌鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经间接ELISA及HI筛选获得了3株能稳定分泌抗HA的MAb杂交瘤细胞,分别命名为1A4、2F5和3D3。MAb亚类鉴定表明,2F5为IgM类,其它为IgG2a亚类,轻链均为κ链。经血凝抑制检测,3株MAb均能与H5亚型的AIV结合,并具有较好的型广谱性。

For the preparation of monoclonal antibodies（MAb） against H5N1 avian influenza virus（AIV）,BALB/c mice were immunized with inactivated H5N1 subtype AIV and the mouse splenic cells were fused with SP2/0 cells.Hybridoma cells stably secreting anti-HA MAbs designated 1A4,2F5,and 3D3,were screened by ELISA coated with the AIV antigen.The immuno-globulin type of MAb identification shown that 2F5 was IgM type with κ chain,others were IgG2a subtype with κ chain.HI test indicated that all the MAbs were reacted against conformational HA epitopes which could be used to identify the H5 subtype AIV.