Purification and Analysis of Strep-Tagged Antibody-Fragments

Part of the Springer Lab Manuals book series (SLM)


The development of generic purification techniques for immunoglobulin (Ig) fragments has gained considerable interest, particularly because the corresponding antigens are often too scarce or unstable to prepare a matrix for affinity chromatography. In this respect, the use of a short peptide tag with defined molecular recognition properties has the advantage that it should not interfere with the function of the antibody fragment and, therefore, its removal is not necessary for most in vitro applications. The Streptag constitutes a nine-amino acid peptide with the sequence “Ala-TrpArg-His-Pro-Gln-Phe-Gly-Gly“, which can easily be fused to scFv, Fv, and Fab fragments. This peptide confers reversible binding activity towards the well known protein-reagent streptavidin. Hence, it enables the purification of a corresponding fusion protein via streptavidin affinity chromatography in one step. Furthermore, the Strep-tag can be used for detection on Western blots or in ELISAs using streptavidin-enzyme conjugates.


Antibody Fragment Host Cell Protein Periplasmic Extract Random Amino Acid Sequence Conserve Restriction Site 
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© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  1. 1.Lehrstuhl für Biologische ChemieTechnische Universität MünchenFreising-WeihenstephanGermany

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