Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.

Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries. Curr Protoc Immunol. 2012 Nov;Chapter 9:Unit9.5 Authors: Pinilla C, Appel JR, Judkowski V, Houghten RA Abstract This unit presents a combinatorial library method that consists of the synthesis and screening of mixture-based synthetic combinatorial libraries of peptide molecules. The protocols employ peptide libraries to identify peptides recognized by MAbs and T cells. The first protocol uses a positional scanning peptide library made up of hexapeptides to identify antigenic determinants recognized by MAbs. The 120 mixtures in the hexapeptide library are tested for their inhibitory activity in a competitive ELISA. The second protocol uses a decapeptide library to identify T cell peptide ligands. The 200 mixtures of the decapeptide library are tested for their ability to induce T cell activation. Support protocols cover optimization of the assay conditions for each MAb or T cell, to achieve the best level of sensitivity and reproducibility, and preparation of a hexapeptide library, along with deconvolution approaches. PMID: 23129156 [PubMed - indexed for MEDLINE]
Source: Current Protocols in Immunology - Category: Allergy & Immunology Tags: Curr Protoc Immunol Source Type: research