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

Taiwanese CDC has decades of experience in antivenom production. In general, in-vivo murine assay was used to access the lethal neutralization titer of antiserum among each time of immunization, and evaluate the capability of antivenom to neutralize the lethal effect of corresponding venoms. However, the ethical consideration for the use of mice in preclinical testing of antivenom has been recently raised. These in-vivo assay will cause considerable suffering and damage the animal welfare for these rodents. In order to follow the rule of 3Rs, the guiding principles for more ethical use of animals in biomedical testing, in this proposal, we plan to develop an ELISA assay to monitor the lethal neutralization titer of antiserum toward N. atra venom during the entire immunization process, which was typically assessed by the in-vivo murine assay. The immunorecognition titers of antiserum against all detectable venom components and venom protein-derived peptides will be systemically evaluated and compared with lethality-neutralizing titer obtained from the in-vivo murine assay, respectively. Venom proteins or venom protein-derived peptides, which show high correlation with the lethality-neutralizing titer, will be selected as the antigens to develop the in-vitro ELISA assays. The potential of using these newly developed assays to replace the in-vivo murine assay in determining the lethality-neutralizing titer of antiserum in the antivenom production process will be thoroughly evaluated.

Project ID:PG10902-0140
External Project ID:MOHW109-CDC-C-114-000114