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Proteomics. 2005 Nov;5(17):4327-37.

Towards a human proteome atlas: high-throughput generation of mono-specific antibodies for tissue profiling.

Author information

1
Department of Biotechnology, Royal Institute of Technology (KTH), Stockholm, Sweden.

Abstract

A great need exists for the systematic generation of specific antibodies to explore the human proteome. Here, we show that antibodies specific to human proteins can be generated in a high-throughput manner involving stringent affinity purification using recombinant protein epitope signature tags (PrESTs) as immunogens and affinity-ligands. The specificity of the generated affinity reagents, here called mono-specific antibodies (msAb), were validated with a novel protein microarray assay. The success rate for 464 antibodies generated towards human proteins was more than 90% as judged by the protein array assay. The antibodies were used for parallel profiling of patient biopsies using tissue microarrays generated from 48 human tissues. Comparative analysis with well-characterized monoclonal antibodies showed identical or similar specificity and expression patterns. The results suggest that a comprehensive atlas containing extensive protein expression and subcellular localization data of the human proteome can be generated in an efficient manner with mono-specific antibodies.

PMID:
16237735
DOI:
10.1002/pmic.200500072
[Indexed for MEDLINE]

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