Comparison of affinity tags for protein purification
Comparison of affinity tags for protein purification
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DOI：
10.1016/j.pep.2005.01.019
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发表时间：
2005-05-01
影响因子：
1.6
通讯作者：
Tan, S
中科院分区：
文献类型：
--
作者：
Lichty, JJ;Malecki, JL;Tan, S
亲和标签是从粗提物中纯化蛋白质的高效工具。为了便于选择亲和标签的纯化项目，我们比较了8个洗脱亲和标签的效率，以纯化蛋白质从大肠杆菌，酵母，果蝇和HeLa提取物。我们的研究结果表明，HIS，CBP，CYD（共价但可解离的NorpD肽），Strep II，FLAG，HPC（蛋白C重链）肽标签，GST和MBP蛋白融合标签系统在纯度，产率和成本上有很大的不同。我们发现，HIS标签提供了良好的收益率的标记蛋白质从廉价的，高容量的树脂，但只有适度的纯度从大肠杆菌提取物和相对较差的纯化从酵母，果蝇，HeLa提取物。CBP标签从大肠杆菌、酵母和果蝇提取物中产生中等纯度的蛋白质，但从HeLa提取物中产生更好的纯度。基于表位的标签，如FLAG和HPC产生的所有提取物的最高纯度的蛋白质，但需要昂贵的，低容量的树脂。我们的研究结果表明，链球菌II标签可以提供一个可接受的妥协，优良的纯化与良好的产量在适度的成本。(c)2005年爱思唯尔公司All rights reserved.
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. (c) 2005 Elsevier Inc. All rights reserved.