New pre-analytical approach for the deep proteome analysis of sera from pancreatitis and pancreas cancer patients.
The presence of high-abundance proteins and the wide dynamic range of protein-distribution complicates the proteome analysis of crude serum. The aim was to establish a new preanalytical protocol for analysis of the deep serum-proteome for biomarker discovery.


METHODS
We investigated the stability and functionality of ProteoMiner™ and tested the new protocol by SELDI-TOF-MS profiling with serum samples obtained from patients with different pancreatic diseases.


RESULTS
We developed a high-throughput protocol and proved the convenience of ProteoMiner™ in the 96-well format to provide insights into the deep serum proteome and facilitate the detection of novel serum biomarkers. Serum samples spiked with defined amounts of insulin, processed with ProteoMiner™ and analyzed by SELDI-TOF-MS revealed that the concentration of the spiked insulin was not altered by ProteoMiner™ treatment.


CONCLUSIONS
ProteoMiner™ technology is robust preanalytical step and can be used in a high-throughput format for analysis of low-abundant proteins in serum.
Fakelman
Frederik
F
Department of General Surgery, University of Heidelberg, Heidelberg, Germany.
Felix
Klaus
K
Büchler
Markus W
MW
Werner
Jens
J
D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000032:analysis / Q000737:chemistry; D001798:Blood Proteins / Q000032:analysis / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D010190:Pancreatic Neoplasms / Q000097:blood / Q000737:chemistry / Q000175:diagnosis; D010195:Pancreatitis / Q000097:blood / Q000175:diagnosis; D040081:Protein Array Analysis / Q000379:methods; D020543:Proteome / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D012107:Research Design; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013057:Spectrum Analysis
D015415:Biomarkers; D001798:Blood Proteins; D020543:Proteome