title: Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format.
abstract: A method for high throughput screening of Green Fluorescent Proteins carrying metal binding tags in bacteria was developed. A random four amino acids tag-peptide library was successfully generated in E. coli. A 96-microtiter plate assembled with metal-iminodiacetic acid small cryogel columns was used for library screening. For the first time we were able to simultaneously screen a metal binding peptide tags library obtained from E. coli against different metal ions. From screening 25 different tags, three clones were able to bind to all metal ions studied (Ni2+, Zn2+, Co2+ and Cd2+). It was clearly demonstrated that the new construct could facilitate the screening of large peptide libraries.
authors.affiliation: Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, PO Box 124, SE-22100 Lund, Sweden.
authors.forename: Amro
authors.initials: A
authors.lastname: Hanora
authors.forename: Florent
authors.initials: F
authors.lastname: Bernaudat
authors.forename: Fatima M
authors.initials: FM
authors.lastname: Plieva
authors.forename: Maria B
authors.initials: MB
authors.lastname: Dainiak
authors.forename: Leif
authors.initials: L
authors.lastname: Bülow
authors.forename: Igor Yu
authors.initials: IY
authors.lastname: Galaev
authors.forename: Bo
authors.initials: B
authors.lastname: Mattiasson
mesh_terms: D000345:Affinity Labels; D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity / Q000295:instrumentation*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D010455:Peptides / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance
chemical_list: D000345:Affinity Labels; D017931:DNA Primers; D010455:Peptides