Wang Qi, Wang Shuai-Yong, Zhang Peng, Liu Xiao-Min, Yu Ling-Xue, Shan Tong-Ling, Tong Wu, Zhou Yan-Jun, Li Guo-Xin, Zheng Hao, Gao Fei, Jiang Yi-Feng, Kong Ning, Li Li-Wei, Tong Guang-Zhi, Yu Hai
1 Shanghai Veterinary Research Institute , Chinese Academy of Agricultural Sciences, Shanghai, China .
2 Shanghai Key Laboratory of Veterinary Biotechnology , Shanghai, China .
Monoclon Antib Immunodiagn Immunother. 2018 Apr;37(2):69-72. doi: 10.1089/mab.2017.0061. Epub 2018 Apr 9.
The purified whole-virus proteins derived from A/swine/Shanghai/1/2014 (H1N1) (SH1) were chosen to immunize BALB/c mice to prepare the monoclonal antibody (MAb) against hemagglutinin (HA) protein of an European avian-like (EA) H1N1 swine influenza virus (SIV). After cloning three times by limiting dilution, one strain of hybridoma cells named 3C7 secreting anti-HA protein MAb was obtained by hybridoma technique. The results of indirect immunofluorescence assay and western blot analyses showed that the MAb 3C7 specifically reacted with the HA protein of EA H1N1 SIV. This work indicated that the MAb 3C7 would be a valuable tool as a specific diagnostic reagent for SIV epidemiological surveys and identification of HA protein epitopes of the EA H1N1 SIVs in the future.
选用源自A/猪/上海/1/2014(H1N1)(SH1)的纯化全病毒蛋白免疫BALB/c小鼠,以制备针对欧洲禽源样(EA)H1N1猪流感病毒(SIV)血凝素(HA)蛋白的单克隆抗体(MAb)。通过有限稀释法克隆三次后,采用杂交瘤技术获得了一株名为3C7的分泌抗HA蛋白MAb的杂交瘤细胞株。间接免疫荧光试验和蛋白质印迹分析结果表明,MAb 3C7能与EA H1N1 SIV的HA蛋白特异性反应。这项工作表明,MAb 3C7作为一种特异性诊断试剂,将成为未来SIV流行病学调查和鉴定EA H1N1 SIVs的HA蛋白表位的宝贵工具。