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.
出处
《中国卫生检验杂志》
CAS
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)