Total apoE levels and the individual apoE isoform (apoE3 and apoE4) levels were previously quantified by use of a mass-spectrometry based method [31 (link)], and the results were reported in a previous pub-lication [21 (link)]. In brief, plasma samples where diluted 1:100, trypsin-digested and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with a Waters NanoAcquity coupled to a Thermo Vantage mass spectrometer at the Mayo Cli-nic Proteomics Core, USA. For quantification of apoE3 and apoE4 isoforms two tryptic peptides, LGADMEDVCGR or LGADMEDVR, derived from the two single major APOE single nucleotide polymorphisms (SNP112 and SNP158) were used. Total apoE was quantified using a peptide present in both isoforms, LGPLVEQGR, which also served as a control to assess the accuracy of the total sum of the two isoforms within each sample.
Protocol detail
Quantifying ApoE Isoforms by LC-MS/MS
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Apoe
Apoe3
Apoe4
Isoforms
Liquid chromatography
Mass spectrometry
Peptide
Plasma
Single nucleotide polymorphisms
Tandem mass spectrometry
Tryptic
Corresponding Organization : Stockholm University
Other organizations : Alzheimer's Association, Arizona State University, University of Arizona, Translational Genomics Research Institute, Mayo Clinic in Arizona
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Variable analysis
independent variables
- None explicitly mentioned
- Total apoE levels
- ApoE3 isoform levels
- ApoE4 isoform levels
- Plasma samples were diluted 1:100
- Trypsin-digestion of plasma samples
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using a Waters NanoAcquity coupled to a Thermo Vantage mass spectrometer
- Peptide LGPLVEQGR used to quantify total apoE as a control to assess the accuracy of the total sum of the two isoforms within each sample
- None explicitly mentioned
- None explicitly mentioned
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