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

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Author

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

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.

Publishing year

2010

Language

English

Pages

2497-2507

Publication/Series

Molecular & Cellular Proteomics

Volume

9

Issue

11

Document type

Journal article

Publisher

American Society for Biochemistry and Molecular Biology

Topic

  • Cancer and Oncology
  • Urology and Nephrology
  • Medicinal Chemistry

Status

Published

Research group

  • Clinical Chemistry, Malmö
  • Urological cancer, Malmö

ISBN/ISSN/Other

  • ISSN: 1535-9484