The AviD-tag, a NeutrAvidin/avidin specific peptide affinity tag for the immobilization and purification of recombinant proteins

Thomas Gaj, Scott C. Meyer, Indraneel Ghosh

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The widespread success of affinity tags throughout the biological sciences has prompted interest in developing new and convenient labeling strategies. Affinity tags are well-established tools for recombinant protein immobilization and purification. More recently these tags have been utilized for selective biological targeting towards multiplexed protein detection in numerous imaging applications as well as for drug-delivery. Recently, we discovered a phage-display selected cyclic peptide motif that was shown to bind selectively to NeutrAvidin and avidin but not to the structurally similar streptavidin. Here, we have exploited this selectivity to develop an affinity tag based on the evolved DRATPY moiety that is orthogonal to known Strep-tag technologies. As proof of principle, the divalent AviD-tag (Avidin-Di-tag) was expressed as a Green Fluorescent Protein variant conjugate and exhibited superior immobilization and elution characteristics to the first generation Strep-tag and a monovalent DRATPY GFP-fusion protein analogue. Additionally, we demonstrate the potential for a peptide based orthogonal labeling strategy involving our divalent AviD-tag in concert with existing streptavidin-based affinity reagents. We believe the AviD-tag and its unique recognition properties will provide researchers with a useful new affinity reagent and tool for a variety of applications in the biological and chemical sciences.

Original languageEnglish (US)
Pages (from-to)54-61
Number of pages8
JournalProtein Expression and Purification
Volume56
Issue number1
DOIs
StatePublished - Nov 2007

Fingerprint

Avidin
Recombinant Proteins
Immobilization
Purification
Peptides
Streptavidin
Biological Science Disciplines
Labeling
Cyclic Peptides
Bacteriophages
Green Fluorescent Proteins
Drug delivery
Proteins
Fusion reactions
Display devices
Research Personnel
Technology
Imaging techniques
neutravidin
Pharmaceutical Preparations

Keywords

  • Affinity
  • AviD-tag
  • Avidin
  • His-tag
  • Immobilization
  • NeutrAvidin streptavidin
  • Orthogonal
  • Purification
  • Strep-tag

ASJC Scopus subject areas

  • Biochemistry

Cite this

The AviD-tag, a NeutrAvidin/avidin specific peptide affinity tag for the immobilization and purification of recombinant proteins. / Gaj, Thomas; Meyer, Scott C.; Ghosh, Indraneel.

In: Protein Expression and Purification, Vol. 56, No. 1, 11.2007, p. 54-61.

Research output: Contribution to journalArticle

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