Tau Protein Quantification in Human Cerebrospinal Fluid by Targeted Mass Spectrometry at High Sequence Coverage Provides Insights into Its Primary Structure Heterogeneity
Abstract
Tau protein plays a major role in neurodegenerative disorders, appears to be a central biomarker of neuronal injury in cerebrospinal fluid (CSF), and is a promising target for Alzheimer's disease immunotherapies. To quantify tau at high sensitivity and gain insights into its naturally occurring structural variations in human CSF, we coupled absolute quantification using protein standard with the multiplex detection capability of targeted high-resolution mass spectrometry (MS) on a Quadrupole-Orbitrap instrument. Using recombinant tau we developed a step-by-step workflow optimization including an extraction protocol that avoided affinity reagents and achieved the monitoring of 22 tau peptides uniformly distributed along the tau sequence. The lower limits of quantification ranged (LLOQ) from 150 to 1500 pg/mL depending on the peptide. Applied to endogenous CSF tau, up to 19 peptides were detected. Interestingly, there were significant differences in the abundance of peptides depending on their position in the sequence, with peptides from the tau mid-domain appearing significantly more abundant than peptides from the N- and C-terminus domains. This MS-based strategy provided results complementary to those of previous ELISA or Western Blot studies of CSF tau and could be applied to tau monitoring in human CSF cohorts.
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Mass spectroscopy studies indicate that CSF tau predominantly consists of tau fragments spanning amino acids 150-250 of the tau protein, with the amino and carboxyl termini presumably having been digested.
“In addition, mass spectroscopy studies indicate that CSF tau predominantly consists of tau fragments spanning amino acids 150–250 of the tau protein, with the amino and carboxyl termini presumably having been digested264,265.”
From Tau-targeting therapies for Alzheimer disease (2018) · cites this paper as [265]
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- Authors
- Barthélemy, N. R., Fenaille, F., Hirtz, C., Sergeant, N., Schraen-Maschke, S., Vialaret, J., Buée, L., Gabelle, A., Junot, C., Lehmann, S., & Becher, F.
- Affiliations
- Inserm (FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- Inserm, UMR-S 1172, Alzheimer & Tauopathies, Centre de Recherche Jean-Pierre Aubert, Lille, Univ. Lille, Faculté de Médecine, Lille F-59045, France
- CEA, iBiTec-S, Service de Pharmacologie et d’Immunoanalyse, Laboratoire d’Etude du Métabolisme des Médicaments, Gif-sur-Yvette 91191, France
- Corresponding author
- Barthélemy, Nicolas R
- ORCID
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-01-19
- Volume
- 15
- Issue
- 2
- Pages
- 667-676
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- OpenAlex
- W2222784460
- Cited by
- 123
- References
- 43
- Citations by year
- 2026: 3 · 2025: 7 · 2024: 15 · 2023: 8 · 2022: 8 · 2021: 19 · 2020: 18 · 2019: 13 · 2018: 17 · 2017: 12 · 2016: 3
- FWCI
- 12.06
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Proteome Research (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Keywords
- tau protein, cerebrospinal fluid, absolute quantification, targeted mass spectrometry, peptide sequence coverage, neurodegenerative diseases, Alzheimer's disease biomarkers
- MeSH terms
- Amino Acid Sequence, Chromatography, Liquid, Humans, Molecular Sequence Data, Peptides, Mass Spectrometry, Reproducibility of Results, Biomarkers, tau Proteins
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