The tandem affinity purification technology: an overview

Biotechnol Lett. 2011 Aug;33(8):1487-99. doi: 10.1007/s10529-011-0592-x. Epub 2011 Mar 22.

Abstract

Tandem affinity purification (TAP) is a methodology for the isolation of protein complexes from endogenous sources. It involves incorporation of a dual-affinity tag into the protein of interest and introduction of the construct into desired cell lines or organisms. Using the two affinity handles, the protein complex assembled under physiological conditions, which contains the tagged target protein and its interacting partners, can be isolated by a sequential purification scheme. Compared with single-step purification, TAP greatly reduces non-specific background and isolates protein complexes with higher purity. TAP-based protein retrieval plus mass spectrometry-based analysis has become a standard approach for identification and characterization of multi-protein complexes. The present article gives an overview of the TAP method, with a focus on its key feature-the dual-affinity tag. In addition, the application of this technology in various systems is briefly discussed.

Publication types

  • Review

MeSH terms

  • Affinity Labels / chemistry*
  • Affinity Labels / metabolism
  • Animals
  • Humans
  • Immunoassay / methods*
  • Mass Spectrometry
  • Protein Binding
  • Recombinant Proteins / isolation & purification*
  • Recombinant Proteins / metabolism

Substances

  • Affinity Labels
  • Recombinant Proteins