Endogeneous peptide profiling of cerebrospinal fluid by MALDI-TOF mass spectrometry: Optimization of magnetic bead-based peptide capture and analysis of preanalytical variables.
Cerebrospinal fluid (CSF) perfuses the brain and spinal cord. CSF contains peptides and proteins important for brain physiology and potentially also relevant to brain pathology. High-throughput endogeneous peptide profiling by MS is an emerging approach for disease diagnosis and biomarker discovery. A magnetic bead-based method for off-line serum peptide capture coupled to MALDI-TOF-MS has been introduced recently. In this study, we optimize the peptide capture method for profiling of CSF and investigate the effect of a number of preanalytical variables. The CSF profiles contain ∼100 reliably detected peptides at m/z 800-4000 with reproducible ion intensities (average 7% CV). The investigated preanalytical variables include: time at room temperature (RT) before storage, storage temperature, freeze-thawing cycles, and blood contamination. The CSF peptidome (<20 kDa) is relatively stable and can withstand a few hours at RT and several freeze-thaw cycles. Several peptides sensitive to storage at -20°C, including Cystatin C, were assigned based on mass or identified by MS/MS. Hemoglobin α and β chains were detected in blood contaminated samples, at levels invisible to the eye (0.01%). These peptides may be used for quality control in a MALDI-TOF-MS screening strategy to select high quality samples for in-depth proteomics analysis in disease studies.
OncoProteomics Laboratory, Department of Medical Oncology, VU Medical Center, Amsterdam, The Netherlands. c.jimenez@vumc.nl.
Connie R
CR
Jimenez
Marleen
M
Koel-Simmelink
Thang V
TV
Pham
Laura
L
van der Voort
Charlotte E
CE
Teunissen