Original Article
Preparation of monoclonal antibodies against the nucleoprotein of influenza A virus and its preliminary applications
Ruxuan Zhang, Enkang Wo, Jianzu Ding, Di Fang, Shuaishuai Chen, Xinyan Yang, Chaotan Guo
Published 2017-04-25
Cite as Inter J Epidemionl Infect Dis, 2017, 44(2): 79-84. DOI: 10.3760/cma.j.issn.1673-4149.2017.02.002
Abstract
ObjectiveTo obtain monoclonal antibodies against nucleoprotein (NP) of influenza A virus, and provide materials for influenza A virus detection kits and related researches.
MethodsThe Balb/c mice were immunized with the purified expression NP of Escherichia coli BL21 (DE3) and their spleen cells were fused with SP2/0 cells. The positive hybridoma cells were selected, and the monoclonal antibodies were obtained by intraperitoneal injection of these positive strains in mice. The purified antibodies were tested by Western blot, ELISA, cell-mediated immunity experiments, double-antibody sandwich ELISA method to confirm the biological characteristic and explore the preliminary applications.
ResultsIt was showed by Western blot, ELISA and cell-mediated immunity experiments that the monoclonal antibodies could specifically combine with both immunogen and NP of H1N1, H3N2 subtypes of influenza virus, and had the influenza virus detection ability. By using double-antibody sandwich ELISA method, it could improve the detection sensitivity up to 128 pg. Moreover, we could observe the location of NP from nuclear to cytoplasm in MDCK cell by using this antibody.
ConclusionsThe prepared NP antibodies can successfully detect H1N1 and H3N2, which are predicted to be the materials of influenza diagnosis kits for flu prevention.
Key words:
Influenza A virus; Nucleoprotein; Monoclonal antibody
Contributor Information
Ruxuan Zhang
Institute of Biological Engineering, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
Enkang Wo
Jianzu Ding
Di Fang
Shuaishuai Chen
Xinyan Yang
Chaotan Guo
