Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.
This unit presents a combinatorial library method that consists of the synthesis and screening of mixture-based synthetic combinatorial libraries (SCL) of peptide molecules. The protocols employ peptide SCL to identify peptides recognized by mAbs and T cells. The first protocol uses a positional scanning SCL (PS-SCL) made up of hexapeptides to identify antigenic determinants recognized by mAbs. The 120 mixtures in the hexapeptide PS-SCL are tested for their inhibitory activity in a competitive ELISA. The second protocol uses a decapeptide PS-SCL to identify T cell peptide ligands. The 200 mixtures of the decapeptide PS-SCL are tested for their ability to induce proliferation. 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 PS-SCL, along with deconvolution approaches.
Pinilla
Clemencia
C
Torrey Pines Institute for Molecular Studies, San Diego, California.
Appel
Jon R
JR
Torrey Pines Institute for Molecular Studies, San Diego, California.
Houghten
Richard A
RA
Torrey Pines Institute for Molecular Studies, San Diego, California.
D000911:Antibodies, Monoclonal / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D019151:Peptide Library
D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D019151:Peptide Library