The FLAG (TM) peptide, a versatile fusion tag for the purification of recombinant proteins

Authors
Einhauer, A Jungbauer, A
Citation
A. Einhauer et A. Jungbauer, The FLAG (TM) peptide, a versatile fusion tag for the purification of recombinant proteins, J BIOCH BIO, 49(1-3), 2001, pp. 455-465
Citations number
55
Categorie Soggetti
Biochemistry & Biophysics
Journal title
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS
ISSN journal
0165022X → ACNP
Volume
49
Issue
1-3
Year of publication
2001
Pages
455 - 465
Database
ISI
SICI code
0165-022X(20011030)49:1-3<455:TF(PAV>2.0.ZU;2-R
Abstract
A fusion tag, called FLAG (TM) and consisting of eight amino acids (AspTyrL ysAspAspAspAspLys) including an enterokinase-cleavage site, was specificall y designed for immunoaffinity chromatography. It allows elution under non-d enaturing conditions [Bio/Technology, 6 (1988) 1204]. Several antibodies ag ainst this peptide have been developed. One antibody, denoted as M1, binds the peptide in the presence of bivalent metal cations, preferably Ca+. Elut ion is effected by chelating agents. Another strategy is competitive elutio n with excess of free FLAG (TM) peptide. Antibodies M2 and M5 are applied i n this procedure. Examples demonstrating the versatility, practicability an d limitations of this technology are given. (C) 2001 Elsevier Science BN. A ll rights reserved.