Affinity proteomics for systematic protein profiling of chromosome 21 gene products in human tissues.
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 toward bioinformatics-designed protein epitope signature tags 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 was 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.
Agaton
Charlotta
C
Department of Biotechnology, AlbaNova University Center, Royal Institute of Technology, Roslagsvägen 30 B, 106 91 Stockholm, Sweden.
Galli
Joakim
J
Höidén Guthenberg
Ingmarie
I
Janzon
Lars
L
Hansson
Marianne
M
Asplund
Anna
A
Brundell
Eva
E
Lindberg
Susanne
S
Ruthberg
Irene
I
Wester
Kenneth
K
Wurtz
Dorothee
D
Höög
Christer
C
Lundeberg
Joakim
J
Ståhl
Stefan
S
Pontén
Fredrik
F
Uhlén
Mathias
M
D000906:Antibodies; D002891:Chromosomes, Human, Pair 21 / Q000378:metabolism*; D019295:Computational Biology; D020224:Expressed Sequence Tags; D006801:Humans; D009928:Organ Specificity; D010865:Pilot Projects; D020543:Proteome / Q000378:metabolism*; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D046888:Tissue Array Analysis
D000906:Antibodies; D020543:Proteome; D011994:Recombinant Proteins