Toward Next Generation Plasma Profiling via Heat-induced Epitope Retrieval and Array-based Assays

Jochen M. Schwenk, Ulrika Igel, Maja Neiman, Hanno Langen, Charlotte Becker, Anders Bjartell, Fredrik Ponten, Fredrik Wiklund, Henrik Gronberg, Peter Nilsson, Mathias Uhlen

Research output: Contribution to journalArticlepeer-review

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. Molecular & Cellular Proteomics 9:2497-2507, 2010.
Original languageEnglish
Pages (from-to)2497-2507
JournalMolecular & Cellular Proteomics
Volume9
Issue number11
DOIs
Publication statusPublished - 2010

Subject classification (UKÄ)

  • Cancer and Oncology
  • Urology and Nephrology
  • Medicinal Chemistry

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