O-028. Immunoproteomic Profiling of Streptococcus pneunomiae Using Human Patient Sera and a Protein Microarray Technology Platform

K. Kwon1, E. Snesrud1, C. Grose1, J. Hasseman1, S. Latham1, D. Yu1, S. Sarria1, T. Mitchell2, G. Pandya1, R. Pieper1, R. Fleischmann1, S. Peterson1;
1J. Craig Venter Inst., Rockville, MD, 2Glasgow Biomedical Res. Cre., Univ. of Glasgow, Glasgow, UNITED KINGDOM.

Protein microarray technologies are increasingly used for high-throughput analysis of protein functions and diagnostic assays. Approximately 700 purified recombinant proteins derived from Streptococcus pneumoniae ORFs cloned into Gateway cassettes were immobilized on protein microarray slides and applied to immunoproteomic analyses. The set of protein consists of hypothetical proteins, extracellular proteins, proteases and house-keeping proteins. First, a small set of proteins was used to optimize technical parameters and the assay methodology for protein microarray analysis with a rabbit anti-S. pneumoniae antibody. Using the optimized immunoproteomic microarray platform, 175 S. pneumoniae sera of patients infected with this pathogen were profiled. This was followed by a multi-dimensional analysis including the assessment of the S. pneumoniae serotype responsible for infection. The proteins identified as highly immunogenic have potential as components of protein-based S. pneumoniae vaccines and are diagnostic or prognostic biomarker candidates. In addition, the data may provide insight into pathways leading to the selective presentation of S. pneumoniae protein epitopes as antigens to the human immune system.