Journal List > J Bacteriol Virol > v.38(4) > 1033910

Kim, Choi, Park, Lee, Kim, Jheong, Kim, Kim, Sohn, and Paik: Purification of Protein Expressed from Three Different Regions of Norovirus (NoV)


Norovirus (NoV), which belongs to the family Caliciviridae, is one of the major causes of nonbacterial acute gastroenteritis in the world. In this study, we purified proteins from the epitope region of norovirus for development of the rapid diagnosis system using polyclonal antibodies. As antigens, parts of the ORF (open reading frame) 2, ORF2-P domain, ORF2-Epi, and ORF3 regions were selected and their expressions were induced. The antigenicity of the purified proteins was identified by Western blotting. Each of the purified proteins was injected into mice for the production of novel antibodies and after 3 months of immunization, sera from the mice were obtained. The polyclonal antibody titer was tested by enzyme-linked immunosorbent assay (ELISA) and antibody against ORF2-Epi showed the highest titer. Those polyclonal antibodies can be used in further immunoassay for the rapid detection of NoVs from food and clinical specimens.


1). Almanza H., Cubillos C., Angulo I., Mateos F., Coastón JR., van der Poel WH., Vinje J., Bárcena J., Mena I. Self-Assembly of the recombinant capsid protein of a swine norovirus into virus-like particles and evaluation of monoclonal antibodies cross-reactive with a human strain from genogroup II. J Clin Microbiol. 46:3971–3979. 2008.
2). Glass RI., Noel J., Ando T., Fankhauser R., Belliot G., Mounts A., Parashar UD., Bresee JS., Monroe SS. The epidemiology of enteric caliciviruses from humans: a reassessment using new diagnosis. J Infect Dis. 181:S254–S261. 2000.
3). Le Guyader FS., Mittelholzer C., Haugarreau L., Hedlund KO., Alsterlund R., Pommepuy M., Svensson L. Detection of noroviruses in rasberries associated with a gastroenteritis outbreak. Int J Food Microbiol. 97:179–186. 2004.
4). Okame M., Shiota T., Hansman G., Takagi M., Yagyu F., Takanashi S., Phan TG., Shimizu Y., Kohno H., Okitsu S., Ushijima H. Anti-norovirus polyclonal antibody and its potential for development of an antigen-ELISA. J Med Virol. 79:1180–1186. 2007.
5). Shiota T., Okame M., Takanashi S., Khamrin P., Takagi M., Satou K., Masuoka Y., Yagyu F., Shimizu Y., Kohno H., Mizuguchi M., Okitsu S., Ushijima H. Characterization of a broadly reactive monoclonal antibody against norovirus genogroups I and II: recognition of a novel conformational epitope. J Virol. 81:12298–12306. 2007.

Figure 1.
Amplified fragments of ORF2-P (816 bp), ORF2-Epi (351 bp) and ORF3 (747 bp) of the NoV capsid gene by RT-PCR.
Figure 2.
Evaluation of the specificity of purified proteins as antigens by SDS-PAGE (left) and Western blotting (right). (A) ORF2-P (43.1 kDa) (B) ORF2-Epi (26.4 kDa) (C) ORF3 (30.7 kDa)
Figure 3.
Titration of the polyclonal antibodies to the ORF2-P, ORF-2-Epi and ORF3 of the NoV capsid by ELISA
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