Production and immunogenicity of VP2 protein of porcine parvovirus expressed in Pichia pastoris

Abstract

Viral protein 2 (VP2) of porcine parvovirus (PPV) is the major viral structural protein and is responsible for eliciting neutralizing antibodies in immunized animals. In this study, we constructed and characterized a recombinant yeast vector encoding the VP2 protein, designated as pGAPZαA-VP2. The construct was confirmed by restriction enzyme digestion, PCR, and sequencing and then introduced into P. pastoris strain SMD1168 by electroporation. The expressed VP2 protein was analyzed by SDS-PAGE and western blot. Immunization of mice with the VP2 protein elicited a PPV-specific humoral immune response. Notably, a preparation of VP2 protein containing adjuvant induced a much better antibody response than VP2 alone. Clearly, the adjuvant strongly enhanced the immunogenicity of VP2. This study provides a foundation for the application of the VP2 protein in the clinical diagnosis of PPV and in vaccination against PPV in the future.

Generation of Recombinant Porcine Parvovirus Virus-Like Particles inSaccharomyces cerevisiaeand Development of Virus-Specific Monoclonal Antibodies

“…To meet the need for stable recombinant VLPs of PPV, several expression systems were tested as an alternative to baculovirus expression system that is a major source of the antigen for the market [26]. E. coli [23], Lactobacillus casei [21], and recently yeast Pichia pastoris [27] were reported to have been successfully used for producing PPV VP2 protein, but VLP formation in these expression systems has not been confirmed. To our knowledge, our study provides the first evidence of stable recombinant PPV VP2 VLPs not produced in baculovirus expression system.…”

“…VLPs generated in baculovirus system exhibit positive immunoreactivity for PPV and are used in most commercial ELISA tests [26]. Most recently, immunogenic PPV VP2 protein was synthesized in yeast Pichia pastoris [27]…”

“…Regarding costs, yield, and ease of handling, VLP production in yeast represents an alternative to the recombinant baculovirus expression system, which is so far the dominating source of VP2-derived VLPs of parvoviruses [27, 28]. …”

Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

“…VLPs generated in baculovirus system exhibit positive immunoreactivity for PPV and are used in most commercial ELISA tests [26]. Most recently, immunogenic PPV VP2 protein was synthesized in yeast Pichia pastoris [27] The formation of recombinant antigenic human parvovirus capsid protein VLPs in S. cerevisiae has been recently demonstrated [28,29]. Regarding costs, yield, and ease of handling, VLP production in yeast represents an alternative to the recombinant baculovirus expression system, which is so far the dominating source of VP2-derived VLPs of parvoviruses [27,28].…”

Nanobody‑horseradish peroxidase and -EGFP fusions as reagents to detect porcine parvovirus in the immunoassays

“…Thus, PPV infection has caused detrimental consequences in the pig industry, such as economic decline. Although the virus has been classified into four clinical genotypes, there is currently only one serotype of PPV [17]. PPV is a non-encapsulated autonomously replicating virus that belongs to the family Parvovirdae, subfamily Parvovirina, and genus Parvovirus [18].…”