Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format.

J Chromatogr A

Department of Biotechnology, Centre for Chemistry and Chemical Engineering, Lund University, PO Box 124, SE-22100 Lund, Sweden.

Published: September 2005

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.

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http://dx.doi.org/10.1016/j.chroma.2005.04.029DOI Listing

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