DEVELOPMENT OF MONOCLONAL ANTIBODIES TO CHEMOKINES IN THE FERRET MODEL OF RESPIRATORY DISEASE.

This proposal addresses the continued pressing need to develop reagents to better understand and characterize the immune response to respiratory infections and support biomarker and countermeasure development through immunologic analyses in the ferret. Select ferret-specific immunoreagents will be generated and tested in the context of influenza infection, where the ferret is the gold standard of preclinical research. The specific and measurable technical objectives are to generate recombinant proteins to eight ferret-specific cytokines and chemokines, generate monoclonal antibodies to six, and screen commercially available antibodies for potential cross-reactivity to two highly-conserved sequences. The antibodies generated will be characterized for binding kinetics via biolayer interferometry, and specificity via knock-in western analysis and competition assays. Confirmation of binding to the intended native antigens will be determined by Luminex assays using serum and nasal wash samples from infected ferrets. Flow cytometry will also be performed using cell surface marker antibodies in conjunction with the antibodies generated using standard intracellular staining protocols. The completion of this Phase I proposal will position Ampersand Biosciences to be near market release of a newly developed ferret-specific 8-plex cytokine/chemokine Luminex panel that will complement our current 13-plex panel on the market.