Publication:
Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein

3
0

Issued Date

2012

Resource Type

Article

File Type

application/pdf

ISSN

1775103

DOI

10.3354/dao02431

Other identifier(s)

2-s2.0-84861668619

Rights Holder(s)

Scopus

Bibliographic Citation

Diseases of Aquatic Organisms. Vol 98, No.2 (2012), p.121-131

Suggested Citation

Wangman P., Senapin S., Chaivisuthangkura P., Longyant S., Rukpratanporn S., Sithigorngul P. Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein. Diseases of Aquatic Organisms. Vol 98, No.2 (2012), p.121-131. doi:10.3354/dao02431 Retrieved from: https://hdl.handle.net/20.500.14740/7094

Title

Production of monoclonal antibodies specific to Macrobrachium rosenbergii nodavirus using recombinant capsid protein

Author(s)

Wangman P.
 Senapin S.
 Chaivisuthangkura P.
 Longyant S.
 Rukpratanporn S.
 Sithigorngul P.

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.

Subject(s)

Antibody
 Biological production
 Coliform bacterium
 Crustacean
 Detection method
 Disease incidence
 Enzyme activity
 Gene expression
 Immunoassay
 Medicine
 Polymerase chain reaction
 Protein
 Virus
 Crangon crangon
 Decapoda (Crustacea)
 Escherichia coli
 Macrobrachium rosenbergii
 Macrobrachium rosenbergii nodavirus
 Miridae
 Nodaviridae
 Capsid protein
 Monoclonal antibody
 Recombinant protein
 Virus antibody
 Animal
 Animal disease
 Article
 Fish disease
 Genetics
 Immunology
 Metabolism
 Mouse
 Nodavirus
 Palaemonidae
 RNA virus infection
 Sensitivity and specificity
 Test strip
 Virology
 Animals
 Antibodies, Monoclonal
 Antibodies, Viral
 Capsid Proteins
 Fish Diseases
 Mice
 Nodaviridae
 Palaemonidae
 Reagent Strips
 Recombinant Proteins
 RNA Virus Infections
 Sensitivity and Specificity

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URI

https://hdl.handle.net/20.500.14740/7094

Collections

Scopus: Year 1983-2021

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