抗H7N9亚型流感病毒神经氨酸酶单克隆抗体的制备及初步鉴定  

Generation and preliminary characterization of monoclonal antibodies against neuraminidase of H7N9 subtype influenza A virus

作　　者：于悦洋 谢怡然[1] 陈迎珠 孙颖[1] 张恒[1] 鲁健[1] 刘丽琦[1] 王大燕[1] 舒跃龙[1] 秦堃[1] 周剑芳[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所、中国国家流感中心,北京100052 [2]温州医科大学、中国疾病预防控制中心病毒病预防控制所

出　　处：《中华实验和临床病毒学杂志》2017年第1期66-70,共5页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的 制备抗H7N9亚型流感病毒神经氨酸酶（Neuraminidase,NA）单克隆抗体,并对单抗的活性与功能进行初步鉴定.方法 利用免疫的BALB/c小鼠脾细胞与Sp2/0骨髓瘤细胞进行融合,获取杂交瘤细胞株,用间接ELISA方法筛选获得阳性杂交瘤细胞克隆,单克隆化后制备小鼠腹水单抗,纯化获得抗H7 N9亚型流感病毒NA的鼠源单克隆抗体,对抗体活性及功能进行鉴定.结果 获得特异性抗H7 N9亚型流感病毒N9鼠源单克隆抗体1G8、3C4和4E8,这三株抗体识别抗原表位不同,3C4和4E8能抑制神经氨酸酶活性,IC50分别为1.45 μg/ml和8.65 μg/ml.结论 3株单克隆抗体均特异性识别A/Anhui/1/2013（H7N9）的N9蛋白,有较好的N9结合活性或能抑制N9酶活性功能.本研究对新型H7N9禽流感病毒的诊断和防控提供了一个良好的思路.Objective To develop the monoclonal antibody （mAb） against neuraminidase of H7N9 subtype influenza A virus and identify its biological function.Methods Female 8 week-old BALB/c mice were immunized and the splenocytes of the mice were fused with Sp2/0 myeloma cells.Indirect ELISA was used to screen hybridoma and the positive clones were subject to be subcloned.Positive clones were identified and the monoclonal antibodies（mAbs） were obtained by purifying the ascetic fluid of mice injected with the hybridoma.The NA-binding as well as neuraminidase-inhibition activity of these mAbs were determined.Results Three mAbs against neuraminidase of H7N9 subtype influenza A virus,1G8,3C4 and 4E8,were obtained.Thcy dcmonstrated different epitop-recognizing.3C4 and 4E8 exhibited neuraminidase inhibitory activity,with a IC50 of 1.45 μg/ml and 8.65 μg/ml,respectively.Conclusions The results suggested that mAbs specific to neuraminidase of H7N9 subtype influenza A virus were developed,providing an useful tool in control and preventing the novel H7N9 influenza A virus.

关 键 词：流感病毒 神经氨酸酶 单克隆抗体 

分 类 号：R392-33[医药卫生—免疫学]