Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format.
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.
Hanora
Amro
A
Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, PO Box 124, SE-22100 Lund, Sweden.
Bernaudat
Florent
F
Plieva
Fatima M
FM
Dainiak
Maria B
MB
Bülow
Leif
L
Galaev
Igor Yu
IY
Mattiasson
Bo
B
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
D000345:Affinity Labels; D017931:DNA Primers; D010455:Peptides