Department of Biology, Indiana University, Bloomington, IN 47405, USA
Howard Hughes Medical Institute, Indiana University, Bloomington, IN 47405, USA
Curr Protein Pept Sci. 2020;21(8):821-830. doi: 10.2174/1389203721666200606220109.
The affinity tags are unique proteins/peptides that are attached at the N- or C-terminus of the recombinant proteins. These tags help in protein purification. Additionally, some affinity tags also serve a dual purpose as solubility enhancers for challenging protein targets. By applying a combinatorial approach, carefully chosen affinity tags designed in tandem have proven to be very successful in the purification of single proteins or multi-protein complexes. In this mini-review, the key features of the most commonly used affinity tags are discussed. The affinity tags have been classified into two significant categories, epitope tags, and protein/domain tags. The epitope tags are generally small peptides with high affinity towards a chromatography resin. The protein/domain tags often perform double duty as solubility enhancers as well as aid in affinity purification. Finally, protease-based affinity tag removal strategies after purification are discussed.
亲和标签是指连接在重组蛋白 N 端或 C 端的独特蛋白质/肽。这些标签有助于蛋白质的纯化。此外，一些亲和标签还可以作为挑战性蛋白靶标的可溶性增强剂发挥双重作用。通过应用组合方法，精心设计的串联亲和标签已被证明在单蛋白或多蛋白复合物的纯化中非常成功。在这篇迷你综述中，讨论了最常用的亲和标签的关键特征。亲和标签被分为两类，表位标签和蛋白/结构域标签。表位标签通常是与层析树脂具有高亲和力的小肽。蛋白/结构域标签通常具有双重作用，既是可溶性增强剂，也是亲和纯化的辅助剂。最后，还讨论了亲和纯化后基于蛋白酶的亲和标签去除策略。
