Generation and Application of Monoclonal Antibodies Specific for the Envelope Protein pORF132 of Cyprinid Herpesvirus 3

Zhenxing Liu; Le Hao; Yanping Ma; Zhiling Liang; Jiangyao Ma; Hao Ke

doi:10.3147/jsfp.51.15

Research Articles

Generation and Application of Monoclonal Antibodies Specific for the Envelope Protein pORF132 of Cyprinid Herpesvirus 3

Zhenxing Liu, Le Hao, Yanping Ma, Zhiling Liang, Jiangyao Ma, Hao Ke

Author information

Zhenxing Liu
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention
Le Hao
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention
Yanping Ma
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention
Zhiling Liang
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention
Jiangyao Ma
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention
Hao Ke
Institute of Animal Heath, Guangdong Academy of Agricultural Sciences, Guangdong Open Laboratory of Veterinary Public Health, Guangdong Provincial Key Laboratory of Livestock Disease Prevention

Keywords: cyprinid herpesvirus 3, KHV, monoclonal antibody, ORF132, ELISA, prokaryotic expression

JOURNAL FREE ACCESS

2016 Volume 51 Issue 1 Pages 15-22

DOI https://doi.org/10.3147/jsfp.51.15

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Abstract

Cyprinid herpesvirus 3 (CyHV-3), also named koi herpesvirus (KHV), is a lethal pathogen of koi and common carp Cyprinus carpio. In the present study, we described a procedure to generate monoclonal antibodies (mAbs) against the envelope protein pORF132 of CyHV-3 and application of the mAb. Three recombinant plasmids targeting the expression of one complete pORF132 and two truncated pORF132 were constructed, respectively. After induction, a truncated pORF132 was successfully expressed in a soluble form and the purified recombinant protein was used in the preparation of mAbs. Two hybridoma cell clones, 4E11 and 4B11, were obtained and mAb 4E11 reacted to CyHV-3 infected cells and not to CyHV-2 (goldfish hematopoietic necrosis virus) and IcHV-1 (channel catfish virus) infected cells. An antigen-capture ELISA based on the pair of mAb 4E11 and anti-CyHV-3 IgY was primarily constructed. This antigen-capture ELISA could distinguish between the CyHV-3 infected and uninfected fish samples.

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