Development of parallel reaction monitoring assays for cerebrospinal fluid proteins associated with Alzheimer's disease

Clin Chim Acta. 2019 Jul:494:79-93. doi: 10.1016/j.cca.2019.03.243. Epub 2019 Mar 9.

Abstract

Detailed knowledge of protein changes in cerebrospinal fluid (CSF) across healthy and diseased individuals would provide a better understanding of the onset and progression of neurodegenerative disorders. In this study, we selected 20 brain-enriched proteins previously identified in CSF by antibody suspension bead arrays (SBA) to be potentially biomarkers for Alzheimer's disease (AD) and verified these using an orthogonal approach. We examined the same set of 94 CSF samples from patients affected by AD (including preclinical and prodromal), mild cognitive impairment (MCI), non-AD dementia and healthy individuals, which had previously been analyzed by SBA. Twenty-eight parallel reaction monitoring (PRM) assays were developed and 13 of them could be validated for protein quantification. Antibody profiles were verified by PRM. For seven proteins, the antibody profiles were highly correlated with the PRM results (r > 0.7) and GAP43, VCAM1 and PSAP were identified as potential markers of preclinical AD. In conclusion, we demonstrate the usefulness of targeted mass spectrometry as a tool for the orthogonal verification of antibody profiling data, suggesting that these complementary methods can be successfully applied for comprehensive exploration of CSF protein levels in neurodegenerative disorders.

Keywords: AD; Alzheimer's disease; Biomarkers; Cerebrospinal fluid; Parallel reaction monitoring (PRM); Suspension bead array (SBA).

MeSH terms

  • Alzheimer Disease / cerebrospinal fluid*
  • Alzheimer Disease / diagnosis*
  • Alzheimer Disease / immunology
  • Antibodies / immunology
  • Antibody Specificity
  • Biomarkers / analysis
  • Cerebrospinal Fluid Proteins / analysis*
  • Cerebrospinal Fluid Proteins / immunology
  • Cohort Studies
  • Humans
  • Mass Spectrometry
  • Protein Array Analysis*

Substances

  • Antibodies
  • Biomarkers
  • Cerebrospinal Fluid Proteins