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 journalArticlepeer-review

20 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

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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