HIGH-DENSITY TILING PEPTIDE ARRAYS COVERING THE COMPLETE Trypanosoma cruzi PROTEOME FOR THE IDENTIFICATION OF NEW CHAGAS DISEASE ANTIGENS AND EPITOPES

BRACCO L; CARMONA SJ; AGÜERO F

Santa Fe

Congreso; XXVIII Reunión Anual de la Sociedad Argentina de Protozoología y Enfermedades Parasitarias y SIMPOSIO Internacional de Biología Celular y Molecular de la Enfermedad de Chagas; 2016

Sociedad Argentina de Protozoología y Enfermedades Parasitarias (SAP)

The full set of antibody (B-cell) specificitiesassociated with the response to a natural infectionremain largely unexplored. We have developed ahighly-multiplexed discovery platform based onnext-generation high-density peptide microarraysand have demonstrated for the first time itspotential to simultaneously identify and finely maphundreds of B-cell epitopes from a complexnatural human infection (Carmona SJ et al 2015).We will next use this platform to obtain a completemap of the antibody specificities developed duringchronic infections with Trypanosoma cruzi(Chagas Disease). For this we developed abioinformatics pipeline written in R and Perl fordesigning peptide array sets that in concert candisplay all 15mer peptides in a proteome ofinterest. Through optimization of the amount ofoverlap between peptides to obtain thecorresponding tiling representations, and throughreduction of redundancy to minimize the numberof repeated peptides, we were able to obtain acomplete representation of our pathogenproteome while reducing significantly the arrayspace required. Each array sector also containrandom sequences to estimate the array signalbaseline (negative control), as well as positivecontrols (known antigens). In this presentation, wewill briefly summarize the use of the high-densitypeptide arrays for immune profiling of B-cellantigens and epitopes and will discuss the newpeptide array designs for whole proteome epitopeprofiling.