PT  - JOURNAL ARTICLE
AU  - Schwenk, Jochen M.
AU  - Igel, Ulrika
AU  - Neiman, Maja
AU  - Langen, Hanno
AU  - Becker, Charlotte
AU  - Bjartell, Anders
AU  - Ponten, Fredrik
AU  - Wiklund, Fredrik
AU  - Grönberg, Henrik
AU  - Nilsson, Peter
AU  - Uhlen, Mathias
TI  - Toward Next Generation Plasma Profiling via Heat-induced Epitope Retrieval and Array-based Assays
AID  - 10.1074/mcp.M110.001560
DP  - 2010 Nov 01
TA  - Molecular & Cellular Proteomics
PG  - 2497--2507
VI  - 9
IP  - 11
4099  - http://www.mcponline.org/content/9/11/2497.short
4100  - http://www.mcponline.org/content/9/11/2497.full
SO  - Mol Cell Proteomics2010 Nov 01; 9
AB  - 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.