Please use this identifier to cite or link to this item: http://repository.li.mahidol.ac.th/dspace/handle/123456789/13478
Title: Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein
Authors: Pradit Wangman
Saengchan Senapin
Parin Chaivisuthangkura
Siwaporn Longyant
Sombat Rukpratanporn
Paisarn Sithigorngul
Srinakharinwirot University
Mahidol University
Thailand National Center for Genetic Engineering and Biotechnology
Chulalongkorn University
Keywords: Agricultural and Biological Sciences
Issue Date: 20-Mar-2012
Citation: Diseases of Aquatic Organisms. Vol.98, No.2 (2012), 121-131
Abstract: The gene encoding the capsid protein of Macrobrachium rosenbergii nodavirus (MrNV) was cloned into pGEX-6P-1 expression vector and then transformed into the Escherichia coli strain BL21. After induction, capsid protein-glutathione-S- transferase (GST-MrNV; 64 kDa) was produced. The recombinant protein was separated using SDS-PAGE, excised from the gel, electro-eluted and then used for immunization for monoclonal antibody (MAb) production. Four MAbs specific to the capsid protein were selected and could be used to detect natural MrNV infections in M. rosenbergii by dot blotting, Western blotting and immunohistochemistry without cross-reaction with uninfected shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was 10 fmol μl-1 of the GST-MrNV, as determined using dot blotting. However, the sensitivity of the MAb on dot blotting with homogenate from naturally infected M. rosenbergii was approximately 200-fold lower than that of 1-step RT-PCR. Immunohistochemical analysis using these MAbs with infected shrimp tissues demonstrated staining in the muscles, nerve cord, gill, heart, loose connective tissue and inter-tubular tissue of the hepatopancreas. Although the positive reactions occurred in small focal areas, the immunoreactivity was clearly demonstrated. The MAbs targeted different epitopes of the capsid protein and will be used to develop a simple immunoassay strip test for rapid detection of MrNV. © Inter-Research 2012.
URI: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84861668619&origin=inward
http://repository.li.mahidol.ac.th/dspace/handle/123456789/13478
ISSN: 16161580
01775103
Appears in Collections:Scopus 2011-2015

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