New pre-analytical approach for the deep proteome analysis of sera from pancreatitis and pancreas cancer patients

Arch Physiol Biochem. 2010 Oct-Dec;116(4-5):208-17. doi: 10.3109/13813455.2010.510891. Epub 2010 Sep 3.

Abstract

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.

Conclusion: ProteoMiner™ technology is robust preanalytical step and can be used in a high-throughput format for analysis of low-abundant proteins in serum.

MeSH terms

  • Adult
  • Aged
  • Biomarkers* / analysis
  • Biomarkers* / chemistry
  • Blood Proteins* / analysis
  • Blood Proteins* / chemistry
  • Chemical Fractionation / methods*
  • Chromatography, Ion Exchange / methods*
  • Female
  • High-Throughput Screening Assays / methods*
  • Humans
  • Male
  • Middle Aged
  • Pancreatic Neoplasms* / blood
  • Pancreatic Neoplasms* / chemistry
  • Pancreatic Neoplasms* / diagnosis
  • Pancreatitis* / blood
  • Pancreatitis* / diagnosis
  • Protein Array Analysis* / methods
  • Proteome / analysis
  • Proteome / chemistry
  • Reproducibility of Results
  • Research Design
  • Sensitivity and Specificity
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Spectrum Analysis

Substances

  • Biomarkers
  • Blood Proteins
  • Proteome