Abstract
Antibody-based proteomics provides a powerful approach for the functional study of the human proteome involving the systematic generation of protein-specific affinity reagents. We used this strategy to construct a comprehensive, antibody-based protein atlas for expression and localization profiles in 48 normal human tissues and 20 different cancers. Here we report a new publicly available database containing, in the first version, approximately 400,000 high resolution images corresponding to more than 700 antibodies toward human proteins. Each image has been annotated by a certified pathologist to provide a knowledge base for functional studies and to allow queries about protein profiles in normal and disease tissues. Our results suggest it should be possible to extend this analysis to the majority of all human proteins thus providing a valuable tool for medical and biological research.
Highlights
Antibody-based proteomics provides a powerful approach for the functional study of the human proteome involving the systematic generation of protein-specific affinity reagents
We show that a comprehensive atlas of human protein expression patterns in normal and cancer tissues can be created by combining the methods mentioned above
The Monospecific Antibodies Used in the Protein Atlas—In this study, we generated monospecific antibodies by a strategy where protein epitope signature tags (PrESTs) are used both as antigens for the development of polyclonal antibodies and as affinity ligands for the subsequent affinity purification of the antibodies as described before [8, 9, 11, 12]
Summary
Antibody-based proteomics provides a powerful approach for the functional study of the human proteome involving the systematic generation of protein-specific affinity reagents. We used this strategy to construct a comprehensive, antibody-based protein atlas for expression and localization profiles in 48 normal human tissues and 20 different cancers. The challenge for antibody-based proteomics is to move from a conventional protein-by-protein approach into a high throughput mode to allow chromosome wide analysis [6, 7] Technical challenges involve both the antigen production and the subsequent generation and characterization of the antibodies. Human Protein Atlas for Normal and Cancer Tissues and the subsequent validation of the specificity of these antibodies by protein microarrays
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