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Comparison of affinity tags for protein purification
Jordan J Lichty1, Joshua L Malecki, Heather D Agnew
1Center for Gene Regulation, Department of Biochemistry and Molecular Biology, 108 Althouse Laboratory, The Pennsylvania State University, University Park, PA 16802-1014, USA.
Protein Expression and Purification
|April 2, 2005
Summary
Selecting the right affinity tag is crucial for protein purification. The Strep II tag offers a good balance of high purity, yield, and moderate cost across various biological extracts.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Chemistry
Background:
- Affinity tags are essential for efficient protein purification from complex biological mixtures.
- Choosing the optimal affinity tag impacts purification success, yield, and cost.
Purpose of the Study:
- To compare the purification efficiency of eight distinct elutable affinity tags.
- To evaluate tag performance across diverse expression systems: E. coli, yeast, Drosophila, and HeLa cells.
Main Methods:
- Purification of tagged proteins using eight different affinity tag systems.
- Assessment of purity, yield, and cost-effectiveness of each tag.
- Analysis of tag performance in various cellular extracts.
Main Results:
- HIS tags yield well from inexpensive resins but offer moderate purity, especially in non-E. coli extracts.
- Epitope tags (FLAG, HPC) provide high purity but require costly, low-capacity resins.
- Strep II tags demonstrate a favorable balance of high purity, good yields, and moderate cost.
Conclusions:
- Affinity tag selection requires careful consideration of the specific application and biological source.
- The Strep II tag presents a versatile option, balancing purification efficacy with economic factors for broad applicability.