Comparison of affinity tags for protein purification

Jordan J. Lichty, Joshua L. Malecki, Heather D. Agnew, Daniel J. Michelson-Horowitz, Song Tan

Research output: Contribution to journalArticle

290 Scopus citations

Abstract

Affinity tags are highly efficient tools for purifying proteins from crude extracts. To facilitate the selection of affinity tags for purification projects, we have compared the efficiency of eight elutable affinity tags to purify proteins from Escherichia coli, yeast, Drosophila, and HeLa extracts. Our results show that the HIS, CBP, CYD (covalent yet dissociable NorpD peptide), Strep II, FLAG, HPC (heavy chain of protein C) peptide tags, and the GST and MBP protein fusion tag systems differ substantially in purity, yield, and cost. We find that the HIS tag provides good yields of tagged protein from inexpensive, high capacity resins but with only moderate purity from E. coli extracts and relatively poor purification from yeast, Drosophila, and HeLa extracts. The CBP tag produced moderate purity protein from E. coli, yeast, and Drosophila extracts, but better purity from HeLa extracts. Epitope-based tags such as FLAG and HPC produced the highest purity protein for all extracts but require expensive, low capacity resin. Our results suggest that the Strep II tag may provide an acceptable compromise of excellent purification with good yields at a moderate cost.

Original languageEnglish (US)
Pages (from-to)98-105
Number of pages8
JournalProtein Expression and Purification
Volume41
Issue number1
DOIs
StatePublished - May 2005

All Science Journal Classification (ASJC) codes

  • Biotechnology

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    Lichty, J. J., Malecki, J. L., Agnew, H. D., Michelson-Horowitz, D. J., & Tan, S. (2005). Comparison of affinity tags for protein purification. Protein Expression and Purification, 41(1), 98-105. https://doi.org/10.1016/j.pep.2005.01.019