TY - JOUR
T1 - Comparison of affinity tags for protein purification
AU - Lichty, Jordan J.
AU - Malecki, Joshua L.
AU - Agnew, Heather D.
AU - Michelson-Horowitz, Daniel J.
AU - Tan, Song
N1 - Funding Information:
We are grateful to Charles Esmon for providing samples of the anti-protein C monoclonal antibody resin, to John Sondek for supplying the pPROEX-HTAInaD expression plasmid and sharing unpublished information, and to Craig Cameron for providing the pET22b-DHFR plasmid. We also thank Decha Sermwittayawong and David Gilmour for the yeast and Drosophila extracts. S.T. is a Pew Scholar in the Biomedical Sciences. This work was supported in part by NIH Grant GM060489 to S.T.
PY - 2005/5
Y1 - 2005/5
N2 - 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.
AB - 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.
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U2 - 10.1016/j.pep.2005.01.019
DO - 10.1016/j.pep.2005.01.019
M3 - Article
C2 - 15802226
AN - SCOPUS:15744372778
SN - 1046-5928
VL - 41
SP - 98
EP - 105
JO - Protein Expression and Purification
JF - Protein Expression and Purification
IS - 1
ER -