Development of ELISA assay to measure the lethality-neutralizing titer of Bungarusmulticinctus antiserum in the production process

Taiwan CDC has been using in-vivo murine assay over decades in accessing antiserum titer among immunized horses. However, the necessity of the use of mice in preclinical testing for antivenom has been critiqued recently. In following the rules of 3Rs to maintain the welfare of model animals, we plan to develop an ELISA assay to monitor the titer of Bungarus multicintus antiserum as an alternative protocol to reduce the suffering and damages of those rodents. Firstly, we will use two dimensional HPLC to separate/purify venom proteins, and examine the identity of each purified protein by mass spectrometry. Then, we will introduce the Toxicity Score formula to calculate and quantify the lethality of the isolated protein components. Aiming at those most lethal ones, we will further apply bioinformatics analysis to obtain the 3D structure of these major toxic proteins to predict their potential toxin-related epitopes, and then synthesize peptides derived from these predicted epitopes. At last, we will develop an ELISA for monitoring the horse antiserum titer against Bungarus multicinctus venom using the major toxin proteins and toxic protein-derived peptides. The protocol is expected to be used as a rapid, economical and animal-friendly approach to detect antiserum titer in the antivenom manufacturing process.

Project ID:PG11001-0024
External Project ID:MOHW110-CDC-C-114-000109