High-density tiling peptide arrays for proteome-wide identification of new Chagas Disease antigens and mapping of antibody epitopes

BRACCO L; CARMONA SJ; AGÜERO F

Ciudad Autonoma de Buenos Aires

Conferencia; 4th ISCB Latin America Conference / 7th Argentinian Congress of Bioinformatics; 2016

International Society for Computational Biology (ISCB) / Asociacion Argentina de Bioinformatica y Biologia Computacional (A2B2C)

high-density tiling peptide arrayS for proteome-WIDE identification of new Chagas Disease antigens and mapping of antibody epitopesLeonel Bracco1, Santiago J Carmona1,2, Fernán Agüero11 Instituto de Investigaciones Biotecnológicas ? Instituto Tecnológico de Chascomús, Universidad de San Martín ? Consejo Nacional de Investigaciones Científicas y Técnicas, San Martín, Buenos Aires, Argentina2 Present address: Ludwig Center for Cancer Research - University of Lausanne - Swiss Institute of Bioinformatics, Lausanne, Switzerland.The full set of antibody (B-cell) specificities associated with the response to a natural infection remain largely unexplored. We have developed a highly-multiplexed discovery platform based on next-generation high-density peptide microarrays and have demonstrated for the first time its potential to simultaneously identify and finely map hundreds of B-cell epitopes from a complex natural human infection (Carmona SJ et al 2015). We will next use this platform to obtain a complete map of the antibody specificities developed during chronic infections with Trypanosoma cruzi (Chagas Disease). For this we developed a bioinformatics pipeline written in R and Perl for designing peptide array sets that in concert can display all 15mer peptides in a proteome of interest. Through optimization of the amount of overlap between peptides to obtain the corresponding tiling representations, and through reduction of redundancy to minimize the number of repeated peptides in each array sector, we were able to obtain a complete representation of our pathogen proteome while reducing significantly the array space required. Each array sector also contain random sequences to estimate the array signal baseline (negative control), as well as positive controls (known antigens). In this presentation we will briefly summarize the use of the high-density peptide arrays for immune profiling of B-cell antigens and epitopes and will discuss the new peptide array designs for whole proteome epitope profiling.Funded by: Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT, Argentina), National Institute of Allergy and Infectious Diseases, NIH (NIH-NIAID, USA).