Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification

Terukazu Nogi, Takeshi Sangawa, Sanae Tabata, Masamichi Nagae, Keiko Tamura-Kawakami, Ayako Beppu, Mitsuharu Hattori, Norihisa Yasui, Junichi Takagi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.

Original languageEnglish
Pages (from-to)2120-2126
Number of pages7
JournalProtein Science
Volume17
Issue number12
DOIs
Publication statusPublished - Dec 2008
Externally publishedYes

Fingerprint

Purification
Peptides
Antibodies
Proteins
Organic solvents
Monoclonal Antibodies
Cells
Costs and Cost Analysis
Kinetics
Water
Costs

Keywords

  • Affinity tag
  • F-spondin
  • Fab fragment
  • Monoclonal antibody
  • Purification
  • Reelin
  • ScFv fragment
  • X-ray crystallography

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Nogi, T., Sangawa, T., Tabata, S., Nagae, M., Tamura-Kawakami, K., Beppu, A., ... Takagi, J. (2008). Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification. Protein Science, 17(12), 2120-2126. https://doi.org/10.1110/ps.038299.108

Novel affinity tag system using structurally defined antibody-tag interaction : Application to single-step protein purification. / Nogi, Terukazu; Sangawa, Takeshi; Tabata, Sanae; Nagae, Masamichi; Tamura-Kawakami, Keiko; Beppu, Ayako; Hattori, Mitsuharu; Yasui, Norihisa; Takagi, Junichi.

In: Protein Science, Vol. 17, No. 12, 12.2008, p. 2120-2126.

Research output: Contribution to journalArticle

Nogi, T, Sangawa, T, Tabata, S, Nagae, M, Tamura-Kawakami, K, Beppu, A, Hattori, M, Yasui, N & Takagi, J 2008, 'Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification', Protein Science, vol. 17, no. 12, pp. 2120-2126. https://doi.org/10.1110/ps.038299.108
Nogi, Terukazu ; Sangawa, Takeshi ; Tabata, Sanae ; Nagae, Masamichi ; Tamura-Kawakami, Keiko ; Beppu, Ayako ; Hattori, Mitsuharu ; Yasui, Norihisa ; Takagi, Junichi. / Novel affinity tag system using structurally defined antibody-tag interaction : Application to single-step protein purification. In: Protein Science. 2008 ; Vol. 17, No. 12. pp. 2120-2126.
@article{ff034d4a6b32492d8dcf8dc5919237cf,
title = "Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification",
abstract = "Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.",
keywords = "Affinity tag, F-spondin, Fab fragment, Monoclonal antibody, Purification, Reelin, ScFv fragment, X-ray crystallography",
author = "Terukazu Nogi and Takeshi Sangawa and Sanae Tabata and Masamichi Nagae and Keiko Tamura-Kawakami and Ayako Beppu and Mitsuharu Hattori and Norihisa Yasui and Junichi Takagi",
year = "2008",
month = "12",
doi = "10.1110/ps.038299.108",
language = "English",
volume = "17",
pages = "2120--2126",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Cold Spring Harbor Laboratory Press",
number = "12",

}

TY - JOUR

T1 - Novel affinity tag system using structurally defined antibody-tag interaction

T2 - Application to single-step protein purification

AU - Nogi, Terukazu

AU - Sangawa, Takeshi

AU - Tabata, Sanae

AU - Nagae, Masamichi

AU - Tamura-Kawakami, Keiko

AU - Beppu, Ayako

AU - Hattori, Mitsuharu

AU - Yasui, Norihisa

AU - Takagi, Junichi

PY - 2008/12

Y1 - 2008/12

N2 - Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.

AB - Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.

KW - Affinity tag

KW - F-spondin

KW - Fab fragment

KW - Monoclonal antibody

KW - Purification

KW - Reelin

KW - ScFv fragment

KW - X-ray crystallography

UR - http://www.scopus.com/inward/record.url?scp=56749168901&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=56749168901&partnerID=8YFLogxK

U2 - 10.1110/ps.038299.108

DO - 10.1110/ps.038299.108

M3 - Article

C2 - 18787202

AN - SCOPUS:56749168901

VL - 17

SP - 2120

EP - 2126

JO - Protein Science

JF - Protein Science

SN - 0961-8368

IS - 12

ER -