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

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

研究成果: Article

16 引用 (Scopus)

抄録

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.

元の言語English
ページ(範囲)693-700
ページ数8
ジャーナルJournal of biochemistry
145
発行部数5
DOI
出版物ステータスPublished - 2009 5 1
外部発表Yes

Fingerprint

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

これを引用

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

:: Journal of biochemistry, 巻 145, 番号 5, 01.05.2009, p. 693-700.

研究成果: Article

Osada, E, Shimizu, Y, Akbar, BK, Kanamori, T & Ueda, T 2009, 'Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system', Journal of biochemistry, 巻. 145, 番号 5, pp. 693-700. https://doi.org/10.1093/jb/mvp027
Osada, Eriko ; Shimizu, Yoshihiro ; Akbar, Bintang K. ; Kanamori, Takashi ; Ueda, Takuya. / Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system. :: Journal of biochemistry. 2009 ; 巻 145, 番号 5. pp. 693-700.
@article{6982b604a52a42a48741e56dbc5d2be4,
title = "Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system",
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.",
keywords = "Antibody, Cell-free protein synthesis system, Epitope mapping, In vitro selection, Ribosome display",
author = "Eriko Osada and Yoshihiro Shimizu and Akbar, {Bintang K.} and Takashi Kanamori and Takuya Ueda",
year = "2009",
month = "5",
day = "1",
doi = "10.1093/jb/mvp027",
language = "English",
volume = "145",
pages = "693--700",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "5",

}

TY - JOUR

T1 - Epitope mapping using ribosome display in a reconstituted cell-free protein synthesis system

AU - Osada, Eriko

AU - Shimizu, Yoshihiro

AU - Akbar, Bintang K.

AU - Kanamori, Takashi

AU - Ueda, Takuya

PY - 2009/5/1

Y1 - 2009/5/1

N2 - 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.

AB - 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.

KW - Antibody

KW - Cell-free protein synthesis system

KW - Epitope mapping

KW - In vitro selection

KW - Ribosome display

UR - http://www.scopus.com/inward/record.url?scp=66749151526&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=66749151526&partnerID=8YFLogxK

U2 - 10.1093/jb/mvp027

DO - 10.1093/jb/mvp027

M3 - Article

C2 - 19228777

AN - SCOPUS:66749151526

VL - 145

SP - 693

EP - 700

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 5

ER -