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Expression of Major Capsid Protein of Cainine Parvovirus by Yeast (Pichia pastoris) and Efficient Purification using Arginine in Affinity Chromatography

Ying He, Qiumei Shi, Sumin Pan, Cairan Yang,Yanying Zhang and Xinhua Shao

An immunochromatographic (IC) assay was developed for rapid detection of canine parvovirus using the monoclonal antibodies (McAbs) against canine parvovirus (CPV-2). To prepare the McAbs, gene encoding the VP2 protein of CPV-2a was expressed in a Pichia pastoris expression vector pPICZ-A. The recombinant VP2 was similar antigenically function to the native capsid protein as demonstrated by Western blotting using CPV- 2 polyclonal antiserum. McAbs against CPV-2 were produced by fusing myeloma cell line SP2/0 with spleen cells from Balb/C mice immunized with purified recombinant VP2 protein. By ELISA it was shown that the McAbs specifically recognized VP2 epitopes of CPV-2 but not those of other canine viruses such as Canine distemper virus (CDV) or canine adenovirus (CAV). An IC assay developed with the McAbs was suitable for rapid detection of canine parvovirus. Fecal samples (120) from dogs suspected of CPV-2 infection were analyzed by both haemaglutination (HA) assay and the IC assay, and 52 and 53 samples were found positive for CPV-2, respectively. Comparison between the two different assays revealed that IC assay is as sensitive as HA; the sensitivity and specificity for the IC assay is 98.6% and 98.1%, respectively.