Antibody-based Profiling of Cerebrospinal Fluid Within Multiple Sclerosis

Proteomics. 2013 Aug;13(15):2256-67. doi: 10.1002/pmic.201200580. Epub 2013 Jun 25.

Abstract

Antibody suspension bead arrays have proven to enable multiplexed and high-throughput protein profiling in unfractionated plasma and serum samples through a direct labeling approach. We here describe the development and application of an assay for protein profiling of cerebrospinal fluid (CSF). While setting up the assay, systematic intensity differences between sample groups were observed that reflected inherent sample specific total protein amounts. Supplementing the labeling reaction with BSA and IgG diminished these differences without impairing the apparent sensitivity of the assay. We also assessed the effects of heat treatment on the analysis of CSF proteins and applied the assay to profile 43 selected proteins by 101 antibodies in 339 CSF samples from a multiple sclerosis (MS) cohort. Two proteins, GAP43 and SERPINA3 were found to have a discriminating potential with altered intensity levels between sample groups. GAP43 was detected at significantly lower levels in secondary progressive MS compared to early stages of MS and the control group of other neurological diseases. SERPINA3 instead was detected at higher levels in all MS patients compared to controls. The developed assay procedure now offers new possibilities for broad-scale protein profiling of CSF within neurological disorders.

Keywords: Antibody microarrays; Biomarker discovery; Cerebrospinal fluid; Multiplexed proteomics technology; Protein arrays; Proteome profiling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Antibodies, Immobilized / metabolism
  • Biomarkers / cerebrospinal fluid
  • Biomarkers / chemistry
  • Cerebrospinal Fluid Proteins / cerebrospinal fluid
  • Cerebrospinal Fluid Proteins / chemistry
  • Cerebrospinal Fluid Proteins / metabolism
  • Female
  • GAP-43 Protein / blood
  • GAP-43 Protein / cerebrospinal fluid
  • Humans
  • Immunoassay / methods*
  • Limit of Detection
  • Male
  • Middle Aged
  • Multiple Sclerosis / cerebrospinal fluid*
  • Multiple Sclerosis / metabolism
  • Protein Array Analysis / methods*
  • Proteome / analysis
  • Proteomics / methods*
  • Serpins / cerebrospinal fluid

Substances

  • Antibodies, Immobilized
  • Biomarkers
  • Cerebrospinal Fluid Proteins
  • GAP-43 Protein
  • Proteome
  • SERPINA3 protein, human
  • Serpins