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Mol Cell Proteomics. 2010 Nov;9(11):2497-507. doi: 10.1074/mcp.M110.001560. Epub 2010 Aug 3.

Toward next generation plasma profiling via heat-induced epitope retrieval and array-based assays.

Author information

  • 1Science for Life Laboratory, Royal Institute of Technology (KTH), Stockholm, Sweden. jochen.schwenk@scilifelab.se

Abstract

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.

PMID:
20682762
[PubMed - indexed for MEDLINE]
PMCID:
PMC2984230
Free PMC Article

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