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Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid
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Neuroscience
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JoVE Journal Neuroscience
Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid

Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid

DOI:
10.3791/56244-v

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10:23 min

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December 04, 2017

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, , , , , , ,
1Inst. of Neuroscience and Physiology, Dept. of Psychiatry and Neurochemistry,Sahlgrenska Academy at University of Gothenburg, 2Clinical Neurochemistry Laboratory,Sahlgrenska University Hospital, 3Department of Molecular Neuroscience,UCL Institute of Neurology

Chapters

  • 00:05Title
  • 01:12Pre-treatment of Bulk Volume CSF without Quantification
  • 02:24Pre-treatment of Small Volume CSF Samples with Isobaric Labeling
  • 04:00MWCO Filtration, Desalting, and Sample Clean-up by Solid Phase Extraction
  • 06:00Offline High-pH Reverse Phase HPLC Sample Fractionation
  • 07:09LC-MS Analysis
  • 07:47Results: Peptide Identification After LC-MS Analysis
  • 09:13Conclusion

Summary

Automatic Translation

Automatic Translation

A method for mass spectrometric analysis of endogenous peptides in human cerebrospinal fluid (CSF) is presented. By employing molecular weight cut-off filtration, chromatographic pre-fractionation, mass spectrometric analysis and a subsequent combination of peptide identification strategies, it was possible to expand the known CSF peptidome nearly ten-fold compared to previous studies.

Tags

Endopeptidomic AnalysisCerebrospinal FluidSample PreparationLC-MSPeptide Pre-fractionationEndogenous PeptidesBiomarkersNeurodegenerative DisordersMWCO FiltrationHigh-pH Reverse Phase HPLCMass Spectrometry

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