title: Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.
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.
authors.affiliation: Torrey Pines Institute for Molecular Studies, San Diego, California.
authors.forename: Clemencia
authors.initials: C
authors.lastname: Pinilla
authors.affiliation: Torrey Pines Institute for Molecular Studies, San Diego, California.
authors.forename: Jon R
authors.initials: JR
authors.lastname: Appel
authors.affiliation: Torrey Pines Institute for Molecular Studies, San Diego, California.
authors.forename: Valeria
authors.initials: V
authors.lastname: Judkowski
authors.affiliation: Torrey Pines Institute for Molecular Studies, San Diego, California.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Houghten
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping / Q000379:methods* / Q000639:trends; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D019151:Peptide Library; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D010446:Peptide Fragments; D019151:Peptide Library