J Biomed Transl Res.  2016 Sep;17(3):58-64. 10.12729/jbtr.2016.17.3.058.

Production and characterization of VP6-specific monoclonal antibodies against bovine group C rotavirus

• Kim DU
• Jeong SA
• Lee SC
• Cho KO
• Kang SY

• KMID: 2368698
• DOI: http://doi.org/10.12729/jbtr.2016.17.3.058

Abstract

Rotaviruses are enteric pathogens causing acute watery dehydrating diarrhea in humans and animals. The importance of group C rotavirus (GpC-RV) infections has not been established as the studies on the GpC-RV have been hampered by the lack of an in vitro culture system. However, diarrheal diseases associated with GpC-RV have been gradually increasing worldwide. In this study, VP6 gene of bovine GpC-RV Korean isolate was expressed, and monoclonal antibodies (mAbs) against VP6 were produced and characterized. The VP6 gene was cloned and expressed based on a baculovirus expression system. Indirect fluorescence antibody (IFA), polymer chain reaction (PCR), and Western blot assays were used to confirm expression of VP6 gene synthesized by the recombinant baculovirus. Eleven mAbs against VP6 were produced using expressed VP6. Cross-reactivity of the mAbs was assessed with recombinant VP6 proteins from porcine GpC-RV and human GpA-RV, or different serotypes of group A rotavirus strains by IFA test. Some mAbs reacted with intact porcine GpC-RV Cowden strain as well as bovine GpC-RV VP6 recombinant baculoviruses, but not with human and animal GpA-RV strains. The VP6-specific mAbs might be useful to develop immunodiagnostic tests such as rapid diagnostic kit, IFA and enzyme-linked immunosorbent assay (ELISA) for detection of GpC-RV.

Keyword

Monoclonal antibodies (mAbs); bovine group C rotavirus; VP6; Baculovirus expression; ELISA

MeSH Terms

Animals
Antibodies, Monoclonal*
Baculoviridae
Blotting, Western
Clone Cells
Diarrhea
Enzyme-Linked Immunosorbent Assay
Fluorescence
Humans
In Vitro Techniques
Polymers
Rotavirus*
Serogroup
Antibodies, Monoclonal
Polymers