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

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.