Standardized approach to proteome profiling of human serum based on magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
BACKGROUND
Magnetic bead purification for the analysis of low-abundance proteins in body fluids facilitates the identification of potential new biomarkers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The aims of our study were to establish a proteome fractionation technique and to validate a standardized blood sampling, processing, and storage procedure for proteomic pattern analysis.


METHODS
We used magnetic bead separation for proteome profiling of human blood by MALDI-TOF MS (mass range, 1000-10,000 Da) and studied the effects on the quality and reproducibility of the proteome analysis of anticoagulants, blood clotting, time and temperature of sample storage, and the number of freeze-thaw cycles of samples.


RESULTS
The proteome pattern of human serum was characterized by approximately 350 signals in the mass range of 1000-10,000 Da. The proteome profile showed time-dependent dynamic changes before and after centrifugation of the blood samples. Serum mass patterns differed between native samples and samples frozen once. The best reproducibility of proteomic patterns was with a single thawing of frozen serum samples.


CONCLUSIONS
Application of the standardized preanalytical blood sampling and storage procedure in combination with magnetic bead-based fractionation decreases variability of proteome patterns in human serum assessed by MALDI-TOF MS.
Baumann
Sven
S
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital, Leipzig, Germany.
Ceglarek
Uta
U
Fiedler
Georg Martin
GM
Lembcke
Jan
J
Leichtle
Alexander
A
Thiery
Joachim
J
D000328:Adult; D015415:Biomarkers / Q000097:blood; D001800:Blood Specimen Collection; D005260:Female; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008280:Magnetics; D008297:Male; D010316:Particle Size; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results; D044967:Serum; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015415:Biomarkers; D020543:Proteome