J Biomed Res.  2015 Sep;16(3):109-114. 10.12729/jbr.2015.16.3.109.

Expression of human norovirus VP1 gene and VP1-specific monoclonal antibodies

• Kim JW
• Kong YJ
• Kim MS
• Lee HJ
• Lee SW
• Kang SY

• KMID: 2169096
• DOI: http://doi.org/10.12729/jbr.2015.16.3.109

Abstract

Norovirus (NoV) is an etiologic agent of human and animal acute gastroenteritis and is a member of the family Caliciviridae. NoV is classified based on nucleotide sequences of the VP1 gene into at least six genogroups (GI-GVI), among which GI, GII, and GIV are known to infect humans and GII is the most prevalent genogroup. In this study, VP1, the full gene of GII human NoV, was cloned from a human fecal sample and expressed using a baculovirus expression system. Human NoV VP1-specific monoclonal antibodies (MAbs) were produced using expressed recombinant VP1. Expressed VP1 in the recombinant virus was confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test, and Western blot analysis. Eight hybridomas secreting VP1-specific MAbs against human GII NoV were generated and characterized. All of the MAbs produced in this study reacted with human GII NoV VP1-recombinant baculoviruses but not with other non-human calicivirus recombinant baculoviruses. These MAbs reacted specifically with human NoV GII.4-2009 virus-like particles (VLPs), and some MAbs showed cross-reactivity with other GII.4 variant VLPs. Expressed human GII NoV VP1-recombinant protein and MAbs specific to this protein can be used as useful reagents for detecting and characterizing human NoV.

Keyword

norovirus; GII; recombinant protein; VP1; monoclonal antibody

MeSH Terms

Animals
Antibodies, Monoclonal*
Baculoviridae
Base Sequence
Blotting, Western
Caliciviridae
Clone Cells
Fluorescence
Gastroenteritis
Genotype
Humans*
Hybridomas
Indicators and Reagents
Norovirus*
Polymerase Chain Reaction
Antibodies, Monoclonal
Indicators and Reagents