Abstract
Abstract A novel proteome microarray representing the majority of the 4086 Yersinia pestis KIM proteins was produced by high-throughput expression and purification. Rabbit hyper-immune sera were produced against proteomes extracted from several pathogenic gram-negative bacteria for use in validation assays. The antibody profile from each of the rabbits enabled detection of: shared cross-reactive proteinssignature proteins common for two or more bacteria, andfingerprint proteins specific to each pathogen. Unique proteins were recognized by convalescence sera from mice that survived plague following immunization with the experimental F1-V vaccine. Several new antigens were discovered that were recognized by antibody from rhesus that survived plague, whereas these Y. pestis proteins were not recognized by sera from animals surviving challenge with spores of the gram-positive Bacillus anthracis. Finally, analysis of sera from cynomolgus macaques acutely infected with Y. pestis or Bacillus anthracis produced antibody-binding patterns that were unique biomarkers for each disease. Our results demonstrate that the whole proteome microarray platform is very useful for establishing diagnostic biomarkers, detecting antibody responses to proteins that could potentially show promise in vaccine development, and also for determining the degree of antigen-cross reactivity between different strains or species of bacteria.
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