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Updated: Jul 18, 2026
A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status
Published on: October 20, 2016
An LC-MS/MS-based platform for the quantification of multiple amyloid beta peptides in surrogate cerebrospinal fluid
Merve Oztug1, Bilgin Vatansever1, Gonca Altin1
1TUBITAK National Metrology Institute (TUBITAK UME), Kocaeli, Turkey.
Introduction:
The accurate quantification of amyloid beta (Aβ) peptides in cerebrospinal fluid (CSF) is crucial for Alzheimer's disease (AD) research, particularly in terms of preclinical and biomarker studies. Traditional methods, such as the enzyme-linked immunosorbent assay (ELISA), have limitations. These include high costs, labor intensity, lengthy processes, and the possibility of cross-reactivity.
Objectives:
The primary objectives of this research were twofold: to comprehensively characterize Aβ peptides and to develop a reliable and accurate method for the simultaneous quantification of Aβ 1-40 and Aβ 1-42 peptides in surrogate CSF that is traceable to the International System of Units (SI).
Methods:
We developed a novel method that combined solid phase extraction (SPE) with isotope dilution liquid chromatography/tandem mass spectrometry (ID-LC/MSMS). SPE was employed to efficiently eliminate matrix interferences, while [15N] Aβ1-40 and [15N] Aβ1-42 served as internal standards to improve accuracy. In addition, we introduced Peptide Impurity Corrected Amino Acid Analysis (PICAA) to ensure traceability to the SI and reliable quantification of Aβ peptides.
Results:
The developed platform demonstrated a linear calibration range of 300-20000 pg/ml for both Aβ1-42 and Aβ1-40 peptides, accompanied by strong correlation coefficients greater than 0.995. Quality Control (QC) samples demonstrated an accuracy of at least 90.0 %.
Conclusion:
The enhanced specificity and flexibility of the developed platform potentially have implications for Alzheimer's disease diagnosis and future investigations of novel Aβ peptide biomarkers.
