H10N8禽流感病毒血凝素中和抗体的结合位点鉴定  

Identification of Epitopes for Neutralizing Antibodies Against H10N8 Avian Influenza Virus Hemagglutinin

作　　者：胡金芳[1] 孙春昀 饶木顶 谢良志[1] 

机构地区：[1]中国医学科学院北京协和医学院细胞工程中心,北京100005 [2]神州细胞工程有限公司新药研发部,北京100176 [3]北京义翘神州生物技术有限公司生物分子研发部,北京100176

出　　处：《中国医学科学院学报》2016年第4期404-410,共7页Acta Academiae Medicinae Sinicae

基　　金：十二五"重大新药创制"科技重大专项(20142X09102044)~~

摘　　要：目的制备并筛选H10N8型禽流感病毒血凝素蛋白的中和抗体,并对其抗原结合位点进行鉴定。方法采用基因工程技术制备H10N8血凝素蛋白单克隆抗体,通过假病毒微量中和实验筛选中和抗体,ELISA和Western blot实验确定抗体结合血凝素蛋白的区域,Discovery Studio 3.5模拟与对接分析软件预测抗原结合位点,进而研究其在H10亚型内的保守性。结果获得1株具有假病毒中和活性的H10N8单克隆抗体,该抗体的结合表位在HA1上,且具有H10亚型内的保守性。结论成功制备1株中和抗体,为H10N8禽流感疫苗和单克隆药物的研究提供了基础。Objective To develop neutralizing monoclonal antibodies（ MAbs） against H10N8 avian influenza virus hemagglutinin and to identify the binding sites. Methods MAbs against hemagglutinin of H10N8 avian influenza virus were developed by genetic engineering. Neutralizing MAbs were screened by microneutralization assay,and then tested by enzyme-linked immunosorbent assay and Western blot to identity the binding sites. The homology modeling process was performed using Discovery Studio 3. 5 software, while the binding epitopes were analyzed by Bio Edit software. Results One MAb that could neutralize the H10N8 pseudovirus was obtained and characterized. Analysis about epitopes suggested that the antibody could bind to the HA1 region of hemagglutinin,while the epitopes on antigen were conserved in H10 subtypes. Conclusions One neutralizing antibody was obtained by this research. The MAb may potentially be further developed as a pre-clinical candidate to treat avian influenza H10N8 virus infection.

关 键 词：H10N8禽流感病毒 血凝素蛋白 中和抗体 表位 

分 类 号：R373.9[医药卫生—病原生物学]