title: Toward next generation plasma profiling via heat-induced epitope retrieval and array-based assays.
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.
authors.affiliation: Science for Life Laboratory, Royal Institute of Technology (KTH), Stockholm, Sweden. jochen.schwenk@scilifelab.se
authors.forename: Jochen M
authors.initials: JM
authors.lastname: Schwenk
authors.forename: Ulrika
authors.initials: U
authors.lastname: Igel
authors.forename: Maja
authors.initials: M
authors.lastname: Neiman
authors.forename: Hanno
authors.initials: H
authors.lastname: Langen
authors.forename: Charlotte
authors.initials: C
authors.lastname: Becker
authors.forename: Anders
authors.initials: A
authors.lastname: Bjartell
authors.forename: Fredrik
authors.initials: F
authors.lastname: Ponten
authors.forename: Fredrik
authors.initials: F
authors.lastname: Wiklund
authors.forename: Henrik
authors.initials: H
authors.lastname: Grönberg
authors.forename: Peter
authors.initials: P
authors.lastname: Nilsson
authors.forename: Mathias
authors.initials: M
authors.lastname: Uhlen
mesh_terms: D000906:Antibodies / Q000378:metabolism; D015415:Biomarkers; D001798:Blood Proteins / Q000032:analysis*; D000939:Epitopes; D057166:High-Throughput Screening Assays / Q000379:methods*; D006358:Hot Temperature; D006801:Humans; D057230:Limit of Detection; D008297:Male; D011471:Prostatic Neoplasms / Q000097:blood; D040081:Protein Array Analysis / Q000379:methods*
chemical_list: D000906:Antibodies; D015415:Biomarkers; D001798:Blood Proteins; D000939:Epitopes