人高致病性禽流感H5N1病毒M2蛋白单克隆抗体的制备及鉴定  

Preparation and evaluation of monoclonal antibodies specific to M2 protein of highly pathogenic avian influenza H5N1 virus

作　　者：于虹[1] 管洁[1] 赵光宇[1] 孙世惠[1] 寇志华[1] 陈万荣[1] 周育森[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《中国卫生检验杂志》2009年第7期1455-1456,1472,共3页Chinese Journal of Health Laboratory Technology

基　　金：禽流感国家863课题(2006AA02Z406);国家支撑计划课题(2006BA06A15);国家重大传染病专项课题(2009ZX10004-401)

摘　　要：目的:以人高致病性禽流感H5N1病毒M2蛋白为靶抗原,制备针对H5N1病毒M2蛋白的单克隆抗体,并对制备的单克隆抗体进行鉴定。方法:用针对H5N1病毒M2蛋白胞外区设计的四分肽作为免疫原免疫BALB/c小鼠,将免疫后的脾细胞与SP2/0骨髓瘤细胞融合,用间接ELISA方法筛选出阳性细胞,经多次亚克隆筛选出稳定分泌特异性抗体的杂交瘤细胞株,制备单抗腹水并进行抗体效价测定及亚类鉴定。结果:获得10株能稳定分泌抗H5N1病毒M2蛋白单克隆抗体杂交瘤细胞株,抗体效价高,抗原特异性强。结论:成功地获得了针对H5N1病毒M2蛋白的单克隆抗体,为H5N1病毒感染的诊断、治疗及病毒致病机制研究奠定了基础。Objective：To produce monoclonal antibodies （mAbs） specific to M2 protein of highly pathogenic avian influenza HSN1 virus and evaluate these mAbs. Methods：The BALB/e mice were immunized with synthetic tetrabranched multiple antigen peptide based on extracellular domain of M2 protein of HSN1 virus, and fusion of lymphocyte and myeloma SP2/0 cell was carried out by conventional method. Antibody titers and specificities of mAbs were measured by ELISA following production of mAbs. Results：We generated 10 hybridoma cell lines with the stable secretion of H5N1 - M2 specific monoclonal antibody （mAb）. Concluslon：The productions of 10 HSNI -M2 specific mAbs with high antibody titers lays a good foundation to perform further studies on diagnosis, therapy and mechanism for infections of HSN1 virus.

关 键 词：人高致病性禽流感H5N1病毒 M2蛋白 单克隆抗体 

分 类 号：R373[医药卫生—病原生物学]