High-density protein microarray is an established technology for characterizing host antibody profiles against entire pathogen proteomes. As one of the highest throughput technologies for antigen discovery, proteome microarrays are a translational research tool for identification of vaccine candidates and biomarkers of susceptibility or protection from microbial challenge. The application has been expanded in recent years due to increased availability of bacterial genomic sequences for a broader range of species and strain diversity. Panproteome microarrays now allow for fine characterization of antibody specificity and cross-reactivity that may be relevant to vaccine design and biomarker discovery, as well as a fuller understanding of factors underlying themes of bacterial evolution and host-pathogen interactions. In this chapter, we present a workflow for design of panproteome microarrays and demonstrate statistical analysis of panproteomic human antibody responses to bacterial vaccination and challenge. Focus is particularly drawn to the bioinformatics and statistical tools and providing nontrivial, real examples that may help foster hypotheses and rational design of panproteomic studies.
Keywords: Antibody profiling; Antigenic diversity; Differential analysis; Panproteome; Protein microarray.
© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.