Development of Polyclonal Antibodies against Porcine Enterovirus G-PLP 3C Protein for Enhanced Diagnosis

Porcine enterovirus G (EV-G) is a small RNA virus belonging to the Enterovirus genus of the Picornaviridae family. While EV-G infection in pigs is usually asymptomatic, it is often associated with other gastrointestinal viruses that cause diarrhea. Notably, recent years have witnessed the emergence of a naturally recombinant EV-G strain with a PLP gene (EV-G-PLP) in various countries. This study focused on developing specific antibodies against EV-G-PLP to enhance diagnostic capabilities.

To achieve this, we prepared polyclonal antibodies targeting the EV-G-PLP 3C protein. This involved constructing a recombinant plasmid (pET-32a-EV-G-PLP-3C), transforming it into BL21 (DE3) cells, and optimizing protein expression conditions. The purified 3C protein was then used to immunize New Zealand rabbits, yielding polyclonal antibodies.

Our results showed that the 3C protein primarily existed as inclusion bodies with a molecular weight of approximately 38.8 kDa. The generated antibodies exhibited good immunogenicity and reactivity as determined by indirect ELISA, indirect immunofluorescence assay, and Western blot analysis. This study provides a valuable tool for the diagnosis of EV-G-PLP and contributes to the understanding of the biological function of the 3C protein.