Identification of B Cell and T Cell Epitopes Using Synthetic Peptide Combinatorial Libraries Article

Pinilla, C, Giulianotti, MA, Santos, RG et al. (2022). Identification of B Cell and T Cell Epitopes Using Synthetic Peptide Combinatorial Libraries . 2(3), 10.1002/cpz1.378

cited authors

  • Pinilla, C; Giulianotti, MA; Santos, RG; Houghten, RA

authors

abstract

  • This article presents a combinatorial library method that consists of the synthesis and screening of mixture-based synthetic combinatorial libraries of peptide molecules to identify B and T cell epitopes. 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. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Screening peptide library for antibody inhibition. Basic Protocol 2: Screening a peptide library to identify CD4+ Or CD8+ T cell ligands. Support Protocol 1: Optimizing antigen and antibody concentrations for screening assay. Support Protocol 2: Preparing a positional scanning peptide library.

publication date

  • March 1, 2022

Digital Object Identifier (DOI)

volume

  • 2

issue

  • 3