Toward next generation plasma profiling via heat-induced epitope retrieval and array-based assays.
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.
Schwenk
Jochen M
JM
Science for Life Laboratory, Royal Institute of Technology (KTH), Stockholm, Sweden. jochen.schwenk@scilifelab.se
Igel
Ulrika
U
Neiman
Maja
M
Langen
Hanno
H
Becker
Charlotte
C
Bjartell
Anders
A
Ponten
Fredrik
F
Wiklund
Fredrik
F
Grönberg
Henrik
H
Nilsson
Peter
P
Uhlen
Mathias
M
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*
D000906:Antibodies; D015415:Biomarkers; D001798:Blood Proteins; D000939:Epitopes