Schmidt Thomas, Skerra Arne
IBA GmbH, Rudolf-Wissell-Str. 28, 37079, Göttingen, Germany,
Methods Mol Biol. 2015;1286:83-95. doi: 10.1007/978-1-4939-2447-9_8.
The Strep-tag-or its improved version Strep-tagII-is an eight amino acid sequence that can be easily fused or conjugated to any protein or peptide of interest and that was engineered for high affinity toward streptavidin, which otherwise is widely known as a tight biotin-binding reagent. Especially in combination with immobilized Strep-Tactin, a mutant streptavidin specifically optimized toward the Strep-tagII, this system enables the facile one-step affinity purification of various biomolecules, including oligomeric and even membrane proteins. The Strep-tagII/Strep-Tactin interaction shows exquisite specificity, thus allowing efficient separation from host cell proteins, and it can be reversed simply by addition of biotin (or a suitable derivative thereof, such as desthiobiotin). Therefore, this system has become very popular for the highly efficient affinity chromatography under biochemically mild conditions. Here, we describe the purification of Strep-tagged proteins from mammalian cell lysates and cell culture supernatants.
链霉亲和标签（Strep-tag）或其改进版本链霉亲和标签II（Strep-tagII）是一种由八个氨基酸组成的序列，它能够轻松地与任何感兴趣的蛋白质或肽融合或偶联，并且经过设计使其对链霉抗生物素蛋白具有高亲和力，而链霉抗生物素蛋白通常作为一种紧密结合生物素的试剂而广为人知。特别是与固定化的链霉抗生物素蛋白衍生物（Strep-Tactin）相结合时，链霉抗生物素蛋白衍生物是一种针对链霉亲和标签II进行了特异性优化的突变体，该系统能够实现各种生物分子（包括寡聚体甚至膜蛋白）的简便一步亲和纯化。链霉亲和标签II/链霉抗生物素蛋白衍生物的相互作用表现出极高的特异性，从而能够有效地从宿主细胞蛋白中分离出来，并且只需添加生物素（或其合适的衍生物，如脱硫生物素）即可使其逆转。因此，该系统在生物化学温和条件下进行高效亲和色谱分离方面变得非常受欢迎。在此，我们描述了从哺乳动物细胞裂解物和细胞培养上清液中纯化带有链霉亲和标签的蛋白质的方法。
