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J Biochem Biophys Methods. 2001 Oct 30;49(1-3):455-65.

The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins.

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  • 1Institute for Applied Microbiology, University of Agriculture and Forestry, Muthgasse 18, 1190 Vienna, Austria.


A fusion tag, called FLAG and consisting of eight amino acids (AspTyrLysAspAspAspAspLys) including an enterokinase-cleavage site, was specifically designed for immunoaffinity chromatography. It allows elution under non-denaturing conditions [Bio/Technology, 6 (1988) 1204]. Several antibodies against this peptide have been developed. One antibody, denoted as M1, binds the peptide in the presence of bivalent metal cations, preferably Ca(+). Elution is effected by chelating agents. Another strategy is competitive elution with excess of free FLAG peptide. Antibodies M2 and M5 are applied in this procedure. Examples demonstrating the versatility, practicability and limitations of this technology are given.

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