Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.
Article
Pinilla, Clemencia, Appel, Jon R, Houghten, Richard A. (2001). Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.
. Chapter 9 9.5.1-9.5.13. 10.1002/0471142735.im0905s45
Pinilla, Clemencia, Appel, Jon R, Houghten, Richard A. (2001). Identification of B cell and T cell epitopes using synthetic peptide combinatorial libraries.
. Chapter 9 9.5.1-9.5.13. 10.1002/0471142735.im0905s45
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.
