Applications of a peptide ligand for streptavidin: the Strep-tag |
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| Authors: | Skerra A Schmidt T G |
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| Institution: | 1. Lehrstuhl für Biologische Chemie, Technische Universität München, D-85350 Freising-Weihenstephan, Germany;2. Institut für Bioanalytik GmbH, Rudolf-Wissell-Str. 28, D-37079 Göttingen, Germany;1. Centro de Biologia Molecular “Severo Ochoa” (Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid), Madrid, Spain;1. Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen, 361021, Fujian, China;2. The Third Institute of Oceanography of the State Oceanic Administration, Xiamen, 361005, Fujian, China;1. Laboratory of Quality & Safety Risk Assessment for Citrus Products (Chongqing), Ministry of Agriculture, Citrus Research Institute, Southwest University, Chongqing 400712, China;2. National Citrus Engineering Research Center, Chongqing 400712, China;1. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, 11800, Penang, Malaysia;2. Usains Biomics Laboratory Testing Services Sdn. Bhd., Universiti Sains Malaysia, Minden, 11800, Penang, Malaysia |
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| Abstract: | The Strep-tag constitutes a nine amino acid-peptide that binds specifically to streptavidin and occupies the same pocket where biotin is normally complexed. Since the Strep-tag participates in a reversible interaction it can be applied for the efficient purification of corresponding fusion proteins on affinity columns with immobilized streptavidin. Elution of the bound recombinant protein can be effected under mild buffer conditions by competition with biotin or a suitable derivative. In addition, Strep-tag fusion proteins can be easily detected in immunochemical assays, like Western blots or ELISAs, by means of commercially available streptavidin-enzyme conjugates. The Strep-tag/streptavidin system has been systematically optimized over the past years, including the engineering of streptavidin itself. Structural insight into the molecular mimicry between the peptide and biotin was furthermore gained from X-ray crystallographic analysis. As a result the system provides a reliable and versatile tool in recombinant protein chemistry. Exemplary applications of the Strep-tag are discussed in this review. |
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