Proteome profiling of human cerebrospinal fluid: exploring the potential of capillary electrophoresis with surface modified capillaries for analysis of complex biological samples.
Artikel i vetenskaplig tidskrift, 2008
A bottom-up proteomic approach, based on capillary electrophoresis (CE) in combination with matrix- assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-ToF/ToF MS), was used to analyze immunoaffinity depleted human cerebrospinal fluid (CSF) and compare it with a non-depleted sample. After enzymatic digestion and desalting, the tryptic peptides were separated by CE using PolyE-323 modified capillaries and fractionated off-line onto MALDI target plates for further analysis by MALDI-MS and MS/MS. The protein profile of the depleted sample was compared with non depleted CSF. Overall, 85 proteins were identified with 95% significance in both samples. The significance scores for proposed biomarkers, such as amyloid-like protein 1 precursor, could be increased up to 12 times after the depletion. Other proteins, often suggested to be related to neurodegenerative diseases, like amyloid beta A4 protein precursor, superoxide dismutase and apolipoprotein E precursor could only be found in the depleted CSF samples. The effect of a derivatization of tryptic peptides with 2- methoxy-4,5-dihydro-1H-imidazole reagent for protein identification with MS was also employed to increase the number of identified proteins and the sequence coverages. The results presented in this study illustrate the benefit of combining a sample pre-fractionation step and a label's ability to enhance the ionization efficiency with the potential of CE using PolyE-323 modified capillaries in the analysis of complex samples. The straight-forward approach that provides speed and simplicity resulting in high-resolution separations and low sample consumption represents an easily applicable separation technique that can serve as a complement to other currently existing analytical approaches needed in modern proteomic analysis of clinically relevant samples.
Molecular Sequence Data
Spectrometry
chemistry
Protein Array Analysis
Amino Acid Sequence
Cerebrospinal Fluid
Capillary
chemistry
Aged
Biomarkers
Adolescent
methods
methods
Middle Aged
instrumentation
Adult
Proteomics
Humans
Matrix-Assisted Laser Desorption-Ionization
instrumentation
Electrophoresis
cerebrospinal fluid
methods
Mass
Peptides
Tandem Mass Spectrometry
cerebrospinal fluid
Författare
Aida Zuberovic
Jörg Hanrieder
Göteborgs universitet
Ulf Hellman
Jonas Bergquist
Magnus Wetterhall
European Journal of Mass Spectrometry
1469-0667 (ISSN)
Vol. 14 4 249-60Ämneskategorier
Analytisk kemi
Neurovetenskaper
DOI
10.1255/ejms.929
PubMed
18756023