Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system

Eriko Osada, Yoshihiro Shimizu, Bintang K. Akbar, Takashi Kanamori, Takuya Ueda

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Ribosome display is a powerful technology for selecting ligand-binding peptides or proteins. We demonstrate here that the ribosome display using the reconstituted cell-free protein synthesis system can be applied for the epitope mapping of monoclonal antibodies (mAbs). Using this technology, we selected peptides that specifically bind to three mAbs from random peptide library. When selection was performed against the anti-FLAG M2 antibody, selected peptides contained previously characterized consensus epitope, indicating that the methodology can be applied for the epitope mapping. When the selection was carried out against two anti-β-Catenin (anti-β-Cat) mAbs, selected peptides had a homology for the partial peptide sequences of β-Cat. Western blot analysis showed that these putative epitopes had affinity for the corresponding mAbs and β-Cat mutants that lack these regions did not bind to the antibodies, indicating we correctly mapped the epitope for these mAbs. The study shown here provides a way for the quick identification of the epitope of mAbs.

Original languageEnglish
Pages (from-to)693-700
Number of pages8
JournalJournal of biochemistry
Volume145
Issue number5
DOIs
Publication statusPublished - 2009 May 1
Externally publishedYes

Fingerprint

Epitope Mapping
Ribosomes
Epitopes
Display devices
Monoclonal Antibodies
Peptides
Proteins
Cats
Technology
Peptide Library
Catenins
Antibodies
Western Blotting
Ligands

Keywords

  • Antibody
  • Cell-free protein synthesis system
  • Epitope mapping
  • In vitro selection
  • Ribosome display

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system. / Osada, Eriko; Shimizu, Yoshihiro; Akbar, Bintang K.; Kanamori, Takashi; Ueda, Takuya.

In: Journal of biochemistry, Vol. 145, No. 5, 01.05.2009, p. 693-700.

Research output: Contribution to journalArticle

Osada, Eriko ; Shimizu, Yoshihiro ; Akbar, Bintang K. ; Kanamori, Takashi ; Ueda, Takuya. / Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system. In: Journal of biochemistry. 2009 ; Vol. 145, No. 5. pp. 693-700.
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