Towards a human proteome atlas: high-throughput generation of mono-specific antibodies for tissue profiling
2005 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 5, no 17, 4327-4337 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2005. Vol. 5, no 17, 4327-4337 p.
Antibody generation, protein microarray, proteome atlas, tissue microarray, tissue profiling
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:uu:diva-80602DOI: 10.1002/pmic.200500072PubMedID: 16237735OAI: oai:DiVA.org:uu-80602DiVA: diva2:108516
LP, KL och JÖ contributed equally to this work.2007-04-182007-04-182015-10-01Bibliographically approved