Quantification of Amyloid-Beta Peptides

Sample processing and mass spectrometric measurements were previously described^{1 (link),2 (link)}. Briefly, Aβ peptides were immunoprecipitated with a mid-domain antibody, in the presence of isotope-labeled internal standard peptides. Concentrations of peptides and fractional isotope labeling were quantified by LC-MS/MS analysis on a Xevo TQ-S (Waters) or TSQ Vantage (ThermoScientific) mass spectrometer. Isotope-labeled leucine was captured from the blood by cation exchange chromatography. The N-heptafluorobutyryl n-propyl ester derivative was quantified by GCMS (Agilent 6890N GC and Agilent 5973N Mass Selective Detector).

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Elbert D.L., Patterson B.W., Lucey B.P., Benzinger T.L, & Bateman R.J. (2022). Importance of CSF-based Aβ clearance with age in humans increases with declining efficacy of blood-brain barrier/proteolytic pathways. Communications Biology, 5, 98.

Publication 2022

Xevo TQ-S TSQ Vantage Agilent 6890N GC Agilent 5973N Mass Selective Detector

Antibody domain Blood Chromatography Ester Gcms Isotope Leucine Mass spectrometric Ms ms Peptides

Corresponding Organization : The University of Texas at Austin

Other organizations : Washington University in St. Louis, Hope Center for Neurological Disorders

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Variable analysis

independent variables

Immunoprecipitation of Aβ peptides with a mid-domain antibody, in the presence of isotope-labeled internal standard peptides

dependent variables

Concentrations of peptides
Fractional isotope labeling
Isotope-labeled leucine concentration

control variables

Isotope-labeled internal standard peptides
Cation exchange chromatography for capturing isotope-labeled leucine from the blood

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