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A more recent version of this article appeared on June 1, 2003.
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Submitted on March 5, 2003
Revised on June 4, 2003
Accepted on June 9, 2003

Affinity Proteomics for systematic protein profiling of chromosome 21 gene products in human tissues

Charlotta Agaton, Joakim Galli, Ingmarie Höidén Guthenberg, Lars Janzon, Marianne Hansson, Anna Asplund, Eva Brundell, Susanne Lindberg, Irene Ruthberg, Kenneth Wester, Dorothee Wurtz, Christer Höög, Joakim Lundeberg, Stefan Ståhl, Fredrik Pontén, and Mathias Uhlén

Biotechnology, Royal Institute of Technology, Stockholm 106 91

Corresponding Author: mathias.uhlen{at}biotech.kth.se

Here we show that an Affinity Proteomics strategy using affinity purified antibodies raised against recombinant human protein fragments can be used for chromosome-wide protein profiling. The approach is based on affinity reagents raised towards bioinformatics-designed Protein Epitope Signature Tags (PrESTs) corresponding to unique regions of individual gene loci. The genes of human chromosome 21 identified by the genome efforts were investigated and the success rates for de novo cloning, protein production and antibody generation were 85%, 76% and 56%, respectively. Using human tissue arrays, a systematic profiling of protein expression and subcellular localization were undertaken for the putative gene products. The results suggest that this Affinity Proteomics strategy can be used to produce a proteome atlas, describing distribution and expression of proteins in normal tissues as well as in common cancers and other forms of diseased tissues.


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