Development and application of a monoclonal antibody against grouper iridovirus (GIV) major capsid protein

Hong-Yi Lin; Chian-Jiun Liou; Yeong-Hsiang Cheng; Hui-Chen Hsu; Jinn-Chin Yiu; Pinwen Peter Chiou; Yu-Shen Lai

doi:10.1016/j.jviromet.2014.04.013

Development and application of a monoclonal antibody against grouper iridovirus (GIV) major capsid protein

J Virol Methods. 2014 Sep 1:205:31-7. doi: 10.1016/j.jviromet.2014.04.013. Epub 2014 May 9.

Authors

Hong-Yi Lin¹, Chian-Jiun Liou², Yeong-Hsiang Cheng³, Hui-Chen Hsu³, Jinn-Chin Yiu⁴, Pinwen Peter Chiou⁵, Yu-Shen Lai⁶

Affiliations

¹ Department of Horticulture, National Ilan University, Yilan, Taiwan; Department of Biotechnology and Animal Science, National Ilan University, Yilan, Taiwan.
² Department of Nursing, Chang Gung University of Science and Technology, Taoyuan, Taiwan; Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, Taiwan.
³ Department of Biotechnology and Animal Science, National Ilan University, Yilan, Taiwan.
⁴ Department of Horticulture, National Ilan University, Yilan, Taiwan.
⁵ Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan.
⁶ Department of Biotechnology and Animal Science, National Ilan University, Yilan, Taiwan. Electronic address: yslai@niu.edu.tw.

PMID: 24814400
DOI: 10.1016/j.jviromet.2014.04.013

Abstract

The major capsid protein (MCP) is a main structural protein of iridoviruses, and is used as a marker for the identification, differentiation and classification of ranaviruses. In the present study, six monoclonal antibodies (mAbs) against recombinant MCP of grouper iridovirus (GIV) were produced and characterized. All of the six mAbs were of IgG1 isotype. Among the mAbs, GIV-MCP-mAb-21 showed the highest reactivity in ELISA and was used to further characterize the expression of GIV-MCP during viral replication. RT-PCR and Western blot analyses revealed that GIV-MCP is a late gene during GIV infection. By immunofluorescence assay, the presence of GIV-MCP was observed in not only the cytoplasm but also the nucleus of GIV-infected cells, a surprising finding that might indicate additional role of GIV-MCP. In conclusion, the newly established GIV-MCP-mAbs are a valuable tool for GIV diagnostic and future studies on GIV pathogenesis.

Keywords: Grouper; Iridovirus; Major capsid protein; Monoclonal antibody.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Antibodies, Monoclonal / immunology
Antibodies, Viral / immunology*
Base Sequence
Capsid Proteins / immunology*
Cell Line
Cytoplasm / metabolism
Enzyme-Linked Immunosorbent Assay / veterinary
Fish Diseases / diagnosis*
Fish Diseases / virology
Fluorescent Antibody Technique / veterinary
Hybridomas
Immunoglobulin G / immunology
Iridovirus / immunology*
Iridovirus / isolation & purification
Mice
Perciformes
Ranavirus / immunology*
Ranavirus / isolation & purification
Recombinant Proteins
Sequence Alignment / veterinary
Sequence Analysis, DNA / veterinary

Substances

Antibodies, Monoclonal
Antibodies, Viral
Capsid Proteins
Immunoglobulin G
Recombinant Proteins
major capsid protein, Iridovirus