WFRF:(Al Khalili Szigyarto Cristina)
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Towards a human proteome atlas : high-throughput generation of mono-specific antibodies for tissue profiling
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- Nilsson, Peter (author)
- KTH,Proteomik
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- Paavilainen, Linda (author)
- Uppsala universitet,Institutionen för genetik och patologi,Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden
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- Larsson, Karin (author)
- KTH,Skolan för bioteknologi (BIO)
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- (creator_code:org_t)
- 2005-10-20
- 2005
- English.
- In: Proteomics. - : Wiley. - 1615-9853 .- 1615-9861. ; 5:17, s. 4327-4337
- Journal article (peer-reviewed)
Abstract
Subject headings
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- 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.
Subject headings
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Keyword
- Antibody generation
- protein microarray
- proteome atlas
- tissue microarray
- tissue profiling
- Bioengineering
Publication and Content Type
- ref (subject category)
- art (subject category)
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- Nilsson, Peter
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- Larsson, Karin
- Ödling, Jenny
- Sundberg, Mårten
- Andersson, Ann-C ...
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- Kampf, Caroline
- Persson, Anja
- Al-Khalili Szigy ...
- Ottoson, Jenny
- Björling, Erik
- Hober, Sophia
- Wernérus, Henrik
- Wester, Kenneth
- Ponten, Fredrik
- Uhlén, Mathias
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- NATURAL SCIENCES
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- Articles in the publication
- Proteomics
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- Uppsala University
- Royal Institute of Technology
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