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

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


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.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / metabolism
  • Biomarkers
  • Blood Proteins / analysis*
  • Epitopes*
  • High-Throughput Screening Assays / methods*
  • Hot Temperature*
  • Humans
  • Limit of Detection
  • Male
  • Prostatic Neoplasms / blood
  • Protein Array Analysis / methods*


  • Antibodies
  • Biomarkers
  • Blood Proteins
  • Epitopes