Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2001 Nov:Chapter 9:9.5.1-9.5.13.
doi: 10.1002/0471142735.im0905s45.

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

Clemencia Pinilla  1 , Jon R Appel  1 , Richard A Houghten  1
Affiliations

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

Clemencia Pinilla et al. Curr Protoc Immunol. 2001 Nov.

Abstract

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.

PubMed Disclaimer

Similar articles

See all similar articles

References

Literature Cited
    1. Atherton, E. and Sheppard, R.C. 1989. Solid Phase Peptide Synthesis: A Practical Approach. IRL Press, Oxford.
    1. Gisin, B.F. 1972. The monitoring of reactions in solid-phase peptide synthesis with picric acid. Anal. Chim. Acta. 58:248-249.
    1. Hemmer, B., Gran, B., Zhao, Y., Marques, A., Pascal, J., Tzou, A., Kondo, T., Cortese, I., Bielekova, B., Straus, S.E., McFarland, H.F., Houghten, R., Simon, R., Pinilla, C., and Martin, R. 1999. Identification of candidate T- cell epitopes and molecular mimics in chronic Lyme disease. Nature Med. 5:1375-1382.
    1. Hemmer, B., Pinilla, C., Gran, B., Vergelli, M., Ling, N., Conlon, P., McFarland, H.F., Houghten, R. and Martin, R. 2000. Contribution of individual amino acids within MHC molecule or antigenic peptide to TCR ligand potency. J. Immunol. 164:861- 871.
    1. Houghten, R.A. 1985. General method for the rapid solid- phase synthesis of large numbers of peptides: Specificity of antigen-antibody interaction at the level of individual amino acids. Proc. Natl. Acad. Sci. U.S.A. 82:5131-5135.
Key References
    1. Houghten et al., 1999. See above.
    1. Pinilla et al., 1999. See above.

MeSH terms

LinkOut - more resources