【Jounal club】常用标签Tag的比较: 纯度，收率，载量，成本等
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Comparison of affinity tags for protein purification
Affinity tags are highly efficient tools for purifying proteins from crude extracts. To facilitate the selection of Affinity tags for purifiation 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 purifiation 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 purifiation with good yields at a moderate cost.
综合考虑纯度、收率和填料成本等方面，研究结果表明Strep II tag是最优选择。
Affinity tags are highly efficient tools for purifying proteins from crude extracts. To facilitate the selection of Affinity tags for purifiation 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 purifiation 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 purifiation with good yields at a moderate cost.
综合考虑纯度、收率和填料成本等方面，研究结果表明Strep II tag是最优选择。
