J Bacteriol Virol.  2008 Dec;38(4):235-237. 10.4167/jbv.2008.38.4.235.

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

• Kim JY
• Choi JW
• Park SW
• Lee SG
• Kim JM
• Jheong WH
• Kim DW
• Kim JM
• Sohn YS
• Paik SY

• KMID: 1483964
• DOI: http://doi.org/10.4167/jbv.2008.38.4.235

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.

Keyword

Norovirus; Purified protein; Polyclonal antibody; ELISA

MeSH Terms

Animals
Antibodies
Blotting, Western
Caliciviridae
Ecthyma, Contagious
Enzyme-Linked Immunosorbent Assay
Gastroenteritis
Humans
Immunization
Immunoassay
Mice
Norovirus
Proteins
Antibodies
Proteins

Figure

• 

  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.

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  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)

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  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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