亲和标签在重组蛋白表达与纯化中的应用

Progress in the Application of Affinity Tags for the Expression and Purification of Recombinant Proteins

亲和标签融合技术为重组蛋白的纯化提供了一种简单方便的纯化工具,具有结合特异性高、洗脱条件温和、通用性强、纯化倍数高等显著优点。概述了亲和标签对融合蛋白表达的影响,可以提高重组蛋白的产量,增强重组蛋白的可溶性,促进重组蛋白的正确折叠;回顾了在重组蛋白表达与纯化中广泛使用的几种亲和标签,以及近年来相继出现的几种比较新颖的纯化标签;介绍了亲和标签的组合使用策略,His6-MBP组合标签集合了两个标签的优点,串联亲和纯化可以纯化获得生理条件下的蛋白质复合体;展望了亲和标签未来的发展趋势,认为仍需继续开发性能更加优越、纯化效果更加显著的纯化标签系统。

Affinity-tag fusion technology provides a simple and convenient tool for the purification of arbitrary recombinant proteins through genetic engineering, which is obviously characterized with high binding specificity, mild elution conditions employed, high generality and hundred-or even thousand-fold purification. The affinity tags widely used for the expression and purification of recombinant proteins is discussed, together with several novel purification tags in recent years. The positive effects of affinity tags on the fusion proteins are presented, which can increase the yield, enhance the solubility and promote the folding of recombinant proteins. An overview of the tag design that combines different affinity tags is introduced, which His6-MBP combinatorial tag has advantages of the two integral parts and tandem affinity purification can purify protein complexes under physiological conditions. Finally, future outlook is also discussed briefly, and purification tag systems that are characterized with more superior performance and more notable purification efficiency are still needed to be developed.