Research on the construction method and characterization of neutralizing mouse-canine chimeric antibody against canine distemper virus

Abstract

Canine distemper (CD) is an acute infectious disease that poses significant health risks to canines. Neutralizing monoclonal antibody (mAb) therapy has demonstrated substantial efficacy in prevent CDV infection. However, immune rejection reactions prevent the use of mouse-derived mAbs in the prophylactic protection of CD. Based on the previously developed neutralizing mAb 9-7B targeting the CDV-H protein, this study successfully utilized both single and dual expression vector strategies to integrate the variable regions of mouse-derived mAb with the constant regions of canine-derived mAb, yielding M-C-H/L and M-C-L-H mouse-canine chimeric antibodies through the CHO expression system. The SDS-PAGE and WB results indicate that both chimeric antibodies possess the correct antibody structure. Both chimeric antibodies specifically recognized the CDV-H protein and CDV in indirect ELISA and indirect immunofluorescence experiments and their neutralizing effect on CDV was confirmed in neutralization assays. This study constructed mouse-canine chimeric antibodies against CDV using two methods for the first time, which provides significant guidance for research on prophylactic CD antibody-based drugs.