Purification of Protein Expressed from Three Different Regions of Norovirus (NoV)

Jin-Young Kim; Jang Won Choi; Seung-Won Park; Sung-Guen Lee; Jong-Min Kim; Weon-Hwa Jheong; Dong-Wook Kim; Jin-Man Kim; Young-Sun Sohn; Soon-Young Paik

doi:10.4167/jbv.2008.38.4.235

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

Original Article

J Bacteriol Virol 2008;38(4):235-237.

Published online: 18 January 2008

DOI: https://doi.org/10.4167/jbv.2008.38.4.235

Purification of Protein Expressed from Three Different Regions of Norovirus (NoV)

Jin-Young Kim¹, Jang Won Choi², Seung-Won Park³, Sung-Guen Lee³, Jong-Min Kim⁴, Weon-Hwa Jheong⁴, Dong-Wook Kim⁵, Jin-Man Kim⁵, Young-Sun Sohn, Soon-Young Paik^3,^∗

¹Department of Food Science and Technology, Ewha Womans University, Seoul, Korea

²Department of Bioindustry, College of Life and Environment, Daegu University, Daegu, Korea

³Department of Microbiology, College of Medicine, the Catholic University of Korea, Seoul, Korea

⁴Department of Environmental Health Research, National Institute of Environmental Research, Incheon, Korea

⁵Catholic Hematopoietic Stem Cell Transplantation Center, the Catholic University of Korea, Seoul, Korea

^∗Corresponding author: Soon-Young Paik. Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, 137-701, Korea. Phone: +82-2-590-1217, Fax: +82-2-535-6473 e-mail: paik@catholic.ac.kr

^∗∗ This work was supported by a Korea Research Foundation Grant (KRF-2003-E00011), Basic Research Program of the Korea Science & Engineering Foundation (R01-2008-000-20343-0) and the BK21 project (2006-0546-3-7).

Revised 24 November 20089 December 2008 Accepted 10 December 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.

Keywords: Norovirus, Purified protein, Polyclonal antibody, ELISA

REFERENCES

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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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