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Toward Next Generation Plasma Profiling via Heat-induced Epitope Retrieval and Array-based Assays
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2010 (English)In: Molecular & Cellular Proteomics, ISSN 1535-9476, E-ISSN 1535-9484, Vol. 9, no 11, 2497-2507 p.Article in journal (Refereed) Published
Abstract [en]

There is a need for high throughput methods for screening patient samples in the quest for potential biomarkers for diagnostics and patient care. Here, we used a combination of undirected target selection, antibody suspension bead arrays, and heat-induced epitope retrieval to allow for protein profiling of human plasma in a novel and systematic manner. Several antibodies were found to reveal altered protein profiles upon epitope retrieval at elevated temperatures with limits of detection improving into lower ng/ml ranges. In a study based on prostate cancer patients, several proteins with differential profiles were discovered and subsequently validated in an independent cohort. For one of the potential biomarkers, the human carnosine dipeptidase 1 protein (CNDP1), the differences were determined to be related to the glycosylation status of the targeted protein. The study shows a path of pursuit for large scale screening of biobank repositories in a flexible and proteome-wide fashion by utilizing heat-induced epitope retrieval and using an antibody suspension bead array format.

Place, publisher, year, edition, pages
2010. Vol. 9, no 11, 2497-2507 p.
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Medical and Health Sciences
URN: urn:nbn:se:uu:diva-148593DOI: 10.1074/mcp.M110.001560ISI: 000285055900013PubMedID: 20682762OAI: oai:DiVA.org:uu-148593DiVA: diva2:402541
Available from: 2011-03-08 Created: 2011-03-08 Last updated: 2011-03-08Bibliographically approved

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