H5亚型禽流感病毒血凝素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

作　　者：但汉并[1] 刘芳芳[1] 黄红亮[1] 金梅林[1] 

机构地区：[1]华中农业大学动物医学院动物病毒室,湖北武汉430070

出　　处：《动物医学进展》2006年第8期67-69,共3页Progress In Veterinary Medicine

基　　金：国家科技攻关项目(2004BA519A07)

摘　　要：以禽流感病毒A／Chicken／Hubei／327／2004（H5N1）免疫Balb／c小鼠，将免疫鼠脾细胞与SP2／0骨髓瘤细胞融合，用血凝抑制试验筛选细胞培养上清，采用有限稀释法对阳性孔进行克隆，3次克隆后获得7株能稳定分泌抗H5亚型禽流感病毒血凝素单克隆抗体的杂交瘤细胞株，分别命名为1C4，1D4，1E12，2E11，4C12，4G2和5E12。细胞培养上清HI效价为2^4～2^7，腹水HI效价可达2^10～2^18。所有单抗与禽流感H7和H9亚型标准血凝抗原，新城疫病毒和鸡传染性支气管炎病毒无交叉反应。在细胞上的中和试验显示具有较高的中和效价，获得的单克隆抗体可在禽流感流行病学的监测中发挥重要作用。Balb/c mice were immunized with the purified AIV antigens. Splenocytes from the immunized mice were fused with SP2/0 myeloma cells. Positive hybridoma clones were screened by hemagglutination inhibition （HI）, and limited dilution method was performed to subclone the positive clones. After three cycles of subcloning, seven McAbs against the hemagglutinin of H5-AIV were obtained, designated as 1C4, 1D4, 1E12, 2E11, 4C12, 4G2 and 5E12. The HI titers of culture supernatant and ascites were 2^4-2^7 and 2^10-2^18. The seven McAbs had good specificity to H5 subtype of avian influenza virus （H5-AIV）. No cross-reactions were found when H7 amd H9 subtypes of avian influenza virus , Newcastle disease virus （NDV） and Avian infectious bronchitis virus （IBV） were tested. All McAbs had high neutralization titers.

关 键 词：禽流感病毒 血凝素 单克隆抗体 

分 类 号：S852.659.5[农业科学—基础兽医学]