The strep-tag system for one-step affinity purification of proteins from mammalian cell culture

Thomas Schmidt, Arne Skerra

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The Strep-tag—or its improved version Strep-tagII—is an eight amino acid sequence that can be easily fused or conjugated to any protein or peptide of interest and that was engineered for high affinity toward streptavidin, which otherwise is widely known as a tight biotin-binding reagent. Especially in combination with immobilized Strep-Tactin, a mutant streptavidin specifically optimized toward the Strep-tagII, this system enables the facile one-step affinity purification of various biomolecules, including oligomeric and even membrane proteins. The Strep-tagII/Strep-Tactin interaction shows exquisite specificity, thus allowing efficient separation from host cell proteins, and it can be reversed simply by addition of biotin (or a suitable derivative thereof, such as desthiobiotin). Therefore, this system has become very popular for the highly efficient affinity chromatography under biochemically mild conditions. Here, we describe the purification of Strep-tagged proteins from mammalian cell lysates and cell culture supernatants.

Original languageEnglish
Pages (from-to)83-95
Number of pages13
JournalMethods in molecular biology (Clifton, N.J.)
Volume1286
DOIs
StatePublished - 2015

Keywords

  • Magnetic beads
  • Mammalian cell culture
  • Peptide tag
  • Protein isolation
  • Recombinant protein expression
  • Strep-Tactin
  • Strep-tag

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