# Preparation of Monoclonal Antibodies Against the gD Protein of Feline Herpesvirus Type-1 by mRNA Immunization

## TL;DR

This study created monoclonal antibodies against a key protein of feline herpesvirus type-1 using mRNA vaccines, enabling better detection and potential diagnostic tools.

## Contribution

The novel use of mRNA immunization to generate monoclonal antibodies against the FHV-1 gD protein for diagnostic applications.

## Key findings

- Five hybridoma cell lines producing anti-FHV-1 gD monoclonal antibodies were successfully generated.
- mAbs D7 and E4 showed the highest specificity and binding activity against FHV-1.
- The antibodies can be used for rapid diagnostic methods like ELISA and colloidal gold assays.

## Abstract

In this research, we designed and developed an mRNA vaccine targeting the feline herpesvirus type-1 (FHV-1) gD protein to immunize mice. Using hybridoma technology, we successfully generated monoclonal antibodies (mAbs) against the FHV-1 gD protein. The gD protein, essential for viral entry, was chosen due to its high conservation and immunogenicity. We obtained five hybridoma cell lines producing anti-FHV-1 gD protein mAbs, with D7 and E4 showing the highest specificity and binding activity. These antibodies serve as specific tools for FHV-1 detection and provide a basis for developing rapid diagnostic methods like ELISA and colloidal gold assays.

This study aimed to develop monoclonal antibodies (mAbs) against the gD protein of FHV-1 for rapid and specific virus detection. The gD protein, a highly conserved part of the FHV-1 envelope, is crucial for viral entry into host cells, making it an ideal detection target. We immunized BALB/c mice with an mRNA vaccine encoding the gD gene, achieving a serum antibody titer of 1:140,000 after three immunizations. The mice were then boosted with recombinant gD protein. Through cell fusion and multiple subcloning rounds, we obtained five hybridoma cell lines (D7, E4, E9, E10, and E19) that stably secrete anti-gD protein mAbs. Characterization by indirect immunofluorescence and Western blot showed that mAbs D7 and E4 have high specificity and strong binding activity against FHV-1, detectable at 2 μg/mL. These mAbs provide specific tools for FHV-1 detection and a basis for developing rapid diagnostic methods using ELISA, colloidal gold, and other technologies.