Dual-color Proteomic Profiling of Complex Samples with a Microarray of 810 Cancer-related Antibodies*
| Autor: | John P. Neoptolemos, Anette Jacob, Martin Sill, Eithne Costello, Andrea S. Bauer, Christoph Schröder, Tatjana Crnogorac-Jurcevic, Tomasz P. Radon, Jörg D. Hoheisel, Manuela Zucknick, Kurt Fellenberg, Sarah Tonack, Sven Rüffer |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Proteomics
Quality Control Antibody microarray Microarray Proteome Antibodies Neoplasm Protein Array Analysis Color Computational biology Biology Biochemistry Analytical Chemistry Neoplasms Humans Molecular Biology Proteomic Profiling Research Molecular biology Pancreatic Neoplasms Case-Control Studies biology.protein Biological Assay DNA microarray Antibody |
| Zdroj: | Molecular & Cellular Proteomics : MCP |
| ISSN: | 1535-9484 1535-9476 |
| Popis: | Antibody microarrays have the potential to enable comprehensive proteomic analysis of small amounts of sample material. Here, protocols are presented for the production, quality assessment, and reproducible application of antibody microarrays in a two-color mode with an array of 1,800 features, representing 810 antibodies that were directed at 741 cancer-related proteins. In addition to measures of array quality, we implemented indicators for the accuracy and significance of dual-color detection. Dual-color measurements outperform a single-color approach concerning assay reproducibility and discriminative power. In the analysis of serum samples, depletion of high-abundance proteins did not improve technical assay quality. On the contrary, depletion introduced a strong bias in protein representation. In an initial study, we demonstrated the applicability of the protocols to proteins derived from urine samples. We identified differences between urine samples from pancreatic cancer patients and healthy subjects and between sexes. This study demonstrates that biomedically relevant data can be produced. As demonstrated by the thorough quality analysis, the dual-color antibody array approach proved to be competitive with other proteomic techniques and comparable in performance to transcriptional microarray analyses. |
| Databáze: | OpenAIRE |
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