BoviScan: a Phage ImmunoPrecipitation Sequencing library to characterize the antibody response against Mycoplasma bovis.

Mycoplasma bovis is a major cattle pathogen and a global concern due to the limited efficient prophylactic tools and the emergence of antibiotic-resistant strains. Current control strategies rely primarily on surveillance, diagnostic and segregation or culling of infected animals. Despite substantial efforts to develop vaccines against M. bovis, no broadly effective solution is currently available. This situation is partly attributable to important gaps in our understanding of the host humoral immune response following infection, vaccination, or challenge. ELISA and Western blot assays have been widely used to assess M. bovis-specific antibody responses. While robust and easy to implement, these approaches are difficult to scale for hundreds of antigens in large animal cohorts and provide limited information on the specific antigens and epitopes targeted by the antibody response. To address these limitations, we developed BoviScan, a Phage Immunoprecipitation Sequencing (PhIP-Seq) library covering the pan-proteome of 295 M. bovis strains. We describe the design, synthesis, and validation of BoviScan using a panel of known-positive sera. We then applied the library to characterize antibody responses in a small cohort of experimentally immunized cattle and to monitor the longitudinal evolution of the humoral response in a single animal. Our results show that BoviScan enables high-throughput, high-resolution mapping of the antibody responses against M. bovis. This approach could be highly valuable for the study of host-pathogen interactions in mycoplasmas and the development of improved vaccines and serological assays.