[以H5N1流感病毒M1蛋白免疫制备单克隆抗体及其鉴定的研究]
[Study on preparation and identification of monoclonal antibodies immunized with H5N1 influenza virus M1 protein].
作者信息
Chen Ai-jun, Yao Li-hong, Guo Jian-qiang, Xu Yi, Liu Xiao-yu, Jia Run-qing, Zhang Zhi-qing
机构信息
State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Dec;23(6):412-4.
OBJECTIVE
To prepare monoclonal antibodies specific for M1 protein of H5N1 subtype human influenza virus, this work may provide new tool in rapid diagnosis and study of type A influenza virus.
METHODS
BALB/c mice were immunized with purified recombinant H5N1 (A/Anhui/1/2005)/M1 protein expressed in E. coli. Spleen cells of the immunized mice were fused with sp2/0 cells to produce hybridoma cell lines. ELISA was performed to identify the monoclonal antibody against M1 protein of H5N1. Immunofluorescence assay (IFA) were applied to identify the specificity of these antibodies.
RESULTS
Three hybridoma cell lines steadily secreting anti-H5N1/M1 McAb were obtained, and their cross reactivity was confirmed by cross-reaction test and IFA.
CONCLUSION
Monoclonal antibodies immunized with H5N1 subtype influenza virus M1 protein are cross-reactive, which can be used to detect different subtype of influenze virus type A.
目的
制备针对H5N1亚型人禽流感病毒M1蛋白的单克隆抗体，为甲型流感病毒的快速诊断及研究提供新工具。
方法
用纯化的在大肠杆菌中表达的重组H5N1（A/安徽/1/2005）/M1蛋白免疫BALB/c小鼠。将免疫小鼠的脾细胞与sp2/0细胞融合以产生杂交瘤细胞系。采用ELISA法鉴定抗H5N1 M1蛋白的单克隆抗体。应用免疫荧光试验（IFA）鉴定这些抗体的特异性。
结果
获得了3株稳定分泌抗H5N1/M1单克隆抗体的杂交瘤细胞系，通过交叉反应试验和IFA证实了它们的交叉反应性。
结论
用H5N1亚型流感病毒M1蛋白免疫制备的单克隆抗体具有交叉反应性，可用于检测不同亚型的甲型流感病毒。
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