Quantitative Matrix-Assisted Laser Desorption Ionization−Fourier Transform Ion Cyclotron Resonance (MALDI−FT-ICR) Peptide Profiling and Identification of Multiple-Sclerosis-Related Proteins

Abstract

We introduce a matrix-assisted laser desorption ionization-Fourier transform ion cyclotron resonance (MALDI-FT-ICR) method for quantitative peptide profiling, using peak height as a measure for abundance. Relative standard deviations in peak height of peptides spiked over 3 orders of magnitude in concentration were below 10% and allowed for accurate comparisons between multiple sclerosis and controls. Application on a set of 163 cerebrospinal fluid (CSF) samples showed significantly differential abundant peptides, which were subsequently identified into proteins (e.g., chromogranin A, clusterin, and complement C3).

Quantitative Proteomics and Metabolomics Analysis of Normal Human Cerebrospinal Fluid Samples

“…MALDI-FT-ICR MS Analysis-The CSF samples were handled according to the same protocol we reported previously (18), in which the (22) as well as peptides in CSF (18), indicating that quantitative MALDI-FT-ICR methods are readily applicable, which is due to the fact that variation in peak height in MALDI-FT-ICR MS is much more reproducible than in, e.g. MALDI-TOF MS.…”

“…In the sample sets, there is a clear division that can be observed between proteins whose abundances vary highly among individuals and proteins that show a much more limited variation between individuals. Among the proteins that showed limited variation between individual CSF samples were serotransferrin (25% RSD in the original sample set, 18% RSD in the validation sample set and 50% RSD in the experimental sample set), tetranectin (21,18, and 52%, respectively), and gelsolin (24,19, and 58%, respectively). Proteins with high variation between individuals in all three sample sets included cadherin-13 (82% RSD in the original sample set, 60% RSD in the validation sample set and 143% RSD in the experimental sample set), contactin-2 (124%, 80 and 156%, respectively), and haptoglobin (135%, 84 and 182%, respectively).…”

Quantitative Proteomics and Metabolomics Analysis of Normal Human Cerebrospinal Fluid Samples*

“…In the sample sets, there is a clear division that can be observed between proteins whose abundances vary highly among individuals and proteins that show a much more limited variation between individuals. Among the proteins that showed limited variation between individual CSF samples were serotransferrin (25% RSD in the original sample set, 18% RSD in the validation sample set and 50% RSD in the experimental sample set), tetranectin (21,18, and 52%, respectively), and gelsolin (24,19, and 58%, respectively). Proteins with high variation between individuals in all three sample sets included cadherin-13 (82% RSD in the original sample set, 60% RSD in the validation sample set and 143% RSD in the experimental sample set), contactin-2 (124%, 80 and 156%, respectively), and haptoglobin (135%, 84 and 182%, respectively).…”

The Impact of Delayed Storage on the Measured Proteome and Metabolome of Human Cerebrospinal Fluid

“…Standardized sample handling is also important when trying to compare results between different laboratories (12)(13)(14). In the search for molecular biomarkers related to disorders of the central nervous system, CSF is the most promising biofluid because of its close contact to the affected tissue (13,(15)(16)(17)(18)(19)(20).…”