# Calcium Chloride Treatment Enhances Antigen Production in Foot-and-Mouth Disease Vaccines for Serotypes SAT1 and SAT3

## TL;DR

Calcium chloride treatment improves antigen production in foot-and-mouth disease vaccines for SAT1 and SAT3 serotypes, enhancing vaccine efficiency.

## Contribution

A novel method using calcium chloride to boost antigen yield in specific FMD vaccine serotypes is introduced.

## Key findings

- A CaCl2 concentration of 3 mM yielded the highest antigen production.
- Supplementing CaCl2 2 hours before or during viral inoculation was most effective.
- CD-BHK 21 medium with 3 mM CaCl2 significantly improved antigen yield for SAT1 BOT and SAT3 ZIM.

## Abstract

Foot-and-mouth disease (FMD) is a highly contagious viral infection causing acute and severe vesicular lesions in cattle and pigs, which has prompted global vaccination policies. This study presents a technique for enhancing antigen yield in SAT1 BOT and SAT3 ZIM by treatment with calcium chloride (CaCl2). We tested changes in cell viability in BHK-21 suspension cells treated with varying concentrations of CaCl2. The optimal CaCl2 concentration was determined based on antigen yield. The timing of CaCl2 supplementation relative to FMD virus inoculation was tested. Finally, the optimal medium for antigen production was identified. We observed a concentration-dependent decrease in BHK-21 cell viability at >7.5 mM CaCl2. A CaCl2 concentration of 3 mM yielded the most antigens. CaCl2 supplementation relative to FMD virus infection was optimal 2 h before or with viral inoculation. CD-BHK 21 medium supplemented with CaCl2 was the most productive medium. Specifically, SAT1 BOT and SAT3 ZIM showed improved antigen production in CD-BHK 21 medium with 3 mM CaCl2, while Provero-1 and Cellvento BHK-200 media showed no significant enhancement. Overall, CaCl2 supplementation enhanced FMD antigen productivity. This study provides a useful framework for enhancing antigen production efficiently in the FMD vaccine industry.