The FLAG peptide, a versatile fusion tag for the purification of recombinant proteins.
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.
Einhauer
A
A
Institute for Applied Microbiology, University of Agriculture and Forestry, Muthgasse 18, 1190 Vienna, Austria. a.einhauer@iam.boku.ac.at
Jungbauer
A
A
D000906:Antibodies / Q000737:chemistry / Q000302:isolation & purification; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D004798:Enzymes / Q000302:isolation & purification; D000939:Epitopes / Q000737:chemistry; D009842:Oligopeptides; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000302:isolation & purification*
D000906:Antibodies; D002352:Carrier Proteins; D004798:Enzymes; D000939:Epitopes; D009842:Oligopeptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C085706:FLAG peptide