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Research Project: Development of a Differential ELISA for ASV Vaccines

Location: Foreign Animal Disease Research

Project Number: 3022-32000-063-031-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: May 1, 2022
End Date: Apr 30, 2025

African swine fever (ASF) is currently causing outbreaks from Central Europe to southeast Asia, causing devastating losses to the swine industry. Currently, there are only experimental live-attenuated vaccines showing a protective effect against this highly virulent outbreak strain. None of these strains have the ability for a serological DIVA (differentiating vaccinated vs non-vaccinated animals). Preliminary tests using peptide coated plates show reactivity with immune serum from pigs vaccinated with ASFV-G-deltaI177L. Optimization of the DIVA ELISA will allow for increased signal to background ratios and provide a suitable assay for commercialization. In addition to the DIVA ELISA, monoclonal antibodies against I177L for future use to increase ELISA sensitivity. Specific objectives: 1. Develop a DIVA ELISA that recognizes I177L to be the companion ELISA for the ASFV-G-dI177L vaccine. 2. Develop Monoclonal antibodies to I177L.

Objective 1: We will optimize reagent formulation and protocols for ELISA, which includes antigens, coating and blocking buffers for plate preparation, assay diluents for sample preparation, and secondary reporters. Protocols include optimizing incubation times and temperatures during manufacturing, storage, and running the ELISA. Objective 2: We will generate and screen monoclonal antibodies (mAb) by standard methods, which will include all stages of development, from injecting mice with antigens to screening and selecting clones for scaled-up mAb production. These antibodies will be used for future use to increase ELISA sensitivity.