Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Apr 16;6(3):fcae132.
doi: 10.1093/braincomms/fcae132. eCollection 2024.

CSF neurofilament light chain profiling and quantitation in neurological diseases

Claire A Leckey  1   2   3 , John B Coulton  4   5 , Tatiana A Giovannucci  1   3   6 , Yingxin He  4   5 , Aram Aslanyan  1 , Rhiannon Laban  3   6 , Amanda Heslegrave  3   6 , Ivan Doykov  2 , Francesca Ammoscato  7 , Jeremy Chataway  8   9 , Floriana De Angelis  8   9 , Sharmilee Gnanapavan  10 , Lauren M Byrne  6 , Jonathan M Schott  1 , Edward J Wild  6 , Nicolas R Barthelémy  4   5 , Henrik Zetterberg  3   6   11   12   13   14 , Selina Wray  6 , Randall J Bateman  4   5 , Kevin Mills  2 , Ross W Paterson  1   3   15
Affiliations

CSF neurofilament light chain profiling and quantitation in neurological diseases

Claire A Leckey et al. Brain Commun. .

Abstract

Neurofilament light chain is an established marker of neuroaxonal injury that is elevated in CSF and blood across various neurological diseases. It is increasingly used in clinical practice to aid diagnosis and monitor progression and as an outcome measure to assess safety and efficacy of disease-modifying therapies across the clinical translational neuroscience field. Quantitative methods for neurofilament light chain in human biofluids have relied on immunoassays, which have limited capacity to describe the structure of the protein in CSF and how this might vary in different neurodegenerative diseases. In this study, we characterized and quantified neurofilament light chain species in CSF across neurodegenerative and neuroinflammatory diseases and healthy controls using targeted mass spectrometry. We show that the quantitative immunoprecipitation-tandem mass spectrometry method developed in this study strongly correlates to single-molecule array measurements in CSF across the broad spectrum of neurodegenerative diseases and was replicable across mass spectrometry methods and centres. In summary, we have created an accurate and cost-effective assay for measuring a key biomarker in translational neuroscience research and clinical practice, which can be easily multiplexed and translated into clinical laboratories for the screening and monitoring of neurodegenerative disease or acute brain injury.

Keywords: mass spectrometry; neurodegenerative diseases; neurofilament light chain.

PubMed Disclaimer

Conflict of interest statement

H.Z. has served at scientific advisory boards and/or as a consultant for AbbVie, Acumen, Alector, Alzinova, ALZPath, Annexon, Apellis, Artery Therapeutics, AZTherapies, Cognito Therapeutics, CogRx, Denali, Eisai, Nervgen, Novo Nordisk, Optoceutics, Passage Bio, Pinteon Therapeutics, Prothena, Red Abbey Labs, reMYND, Roche, Samumed, Siemens Healthineers, Triplet Therapeutics and Wave, has given lectures in symposia sponsored by Cellectricon, Fujirebio, Alzecure, Biogen and Roche and is a co-founder of Brain Biomarker Solutions in Gothenburg AB (BBS), which is a part of the GU Ventures Incubator Program (outside submitted work). R.J.B. and R.W.P. lead The Neurofilament Light Consortium, an industry academic collaboration that is supported by AbbVie, Bristol Myers Squibb, Biogen and Roche. R.W.P. has received honoraria from GE Healthcare for educational talks, which is used to support academic work. R.J.B. has received research funding from Avid Radiopharmaceuticals, Janssen, Roche/Genentech, Eli Lilly, Eisai, Biogen, AbbVie, Bristol Myers Squibb and Novartis. Washington University and R.J.B. have equity ownership interest in C2N Diagnostics and receive income based on technology (neurofilament light chain assays and materials) licenced by Washington University to C2N Diagnostics. R.J.B. receives income from C2N Diagnostics for serving on the scientific advisory board. R.J.B. serves on the Roche Gantenerumab Steering Committee as an unpaid member. J.C. has received support from the Health Technology Assessment (HTA) Programme (National Institute for Health Research, NIHR), the UK MS Society, the US National MS Society and the Rosetrees Trust. He is supported in part by the National Institute for Health and Care Research University College London Hospitals Biomedical Research Centre, London, UK. He has been a local principal investigator for a trial in multiple sclerosis funded by the Multiple Sclerosis Society of Canada. A local principal investigator for commercial trials was funded by Ionis, Novartis and Roche and has taken part in advisory boards/consultancy for Azadyne, Biogen, Lucid, Janssen, Merck, NervGen, Novartis and Roche. The MS-SMART trial was funded by the Efficacy and Mechanism Evaluation programme as project number 11/30/11. MS-SMART is an investigator-led project sponsored by the University College London. This independent research is awarded by and funded by the Medical Research Council, the UK MS Society and the National MS Society and is managed by the National Institute for Health Research on behalf of the Medical Research Council–National Institute for Health partnership. Additional support came from the University of Edinburgh, the National Institute for Health Research University College London Hospitals Biomedical Research Centre and University College London and the National Institute for Health Research Leeds Clinical Research Facility (Dental Translational and Clinical Research Unit). Riluzole was provided without charge by Sanofi-Genzyme who was not involved in either the trial design, running of the trial or analysis. F.D.A. received speaker honoraria from Neurology Academy, Janssen, Merck, Novartis and Sanofi; is on the advisory board for Novartis and Coloplast; has received congress fees from Janssen, Merck, Novartis and Roche; and is a regional coordinator for the Oratorio Hand Trial (Hoffmann-La Roche). The rest of the authors declared no conflicting interests.

Figures

Graphical Abstract
Graphical Abstract
This image was created with BioRender.com.
Figure 1
Figure 1
NfL profiles in CSF across neurodegenerative disease groups show greatest detection of NfL species from the Coil 2B domain. NfL concentrations determined by IP–MS (WashU) are plotted by the peptide amino acid (AA) residue and represent NfL species located in Coil 1A (NfL93–124), Coil 1B (NfL138–234) and Coil 2B (NfL281–396) of the mid-rod domain and subdomain B (NfL444–543) of the C-terminal tail domain. (A–H) Traces represent individual NfL peptide concentrations, with control or disease states detailed by panel legends. (I) Mean fold change profiles across NfL peptides for each disease state compared to control and (J) mean NfL peptide fold change (compared to controls) for each disease state (horizontal bars represent mean fold change and dots represent fold change for each monitored peptide). Detected peptide sequences by NfL domain and amino acid residues are specified in Supplementary Table 1. Ad, Alzheimer’s disease; bvFTD, behavioural variant frontotemporal dementia; CBS, corticobasal syndrome; DLB, dementia with Lewy bodies; HD, Huntington’s disease; IP–MS, immunoprecipitation–mass spectrometry; MS, multiple sclerosis; NfL, neurofilament light chain; SD, semantic dementia; WashU, Washington University in St Louis.
Figure 2
Figure 2
NfL concentration measured by Simoa strongly correlates with the IP–MS methods for NfL316–323 peptide (TLEIEACR) developed at each study centre. Spearman’s correlations between NfL concentrations as measured by (A) IP–MS/MS (UCL) and Simoa (r = 0.90, P < 0.001), (B) IP–MS (WashU) and Simoa (r = 0.89, P < 0.001) and (C) IP–MS/MS (UCL) and IP-MS (WashU; r = 0.86, P < 0.001) are shown. NfL data plotted for both MS assays represent NfL316–323 (TLEIEACR). Strong correlation is observed across all method comparisons, with the highest correlation observed between IP–MS/MS (UCL) and Simoa methods. Ad, Alzheimer’s disease; bvFTD, behavioural variant frontotemporal dementia; CBS, corticobasal syndrome; DLB, dementia with Lewy bodies; HD, Huntington’s disease; IP–MS/MS, immunoprecipitation–tandem mass spectrometry; IP–MS, immunoprecipitation–mass spectrometry; MS, multiple sclerosis; NfL, neurofilament light chain; SD, semantic dementia; Simoa, single-molecule array; UCL, University College London; WashU, Washington University in St Louis.
Figure 3
Figure 3
Evaluation of method agreement between IP–MS/MS (UCL) and Simoa NfL measures demonstrates good agreement between assays. Bland–Altman analysis to show differences in log-transformed NfL concentrations measured by IP–MS/MS (UCL) and Simoa against the mean log NfL concentration of the methods, with the line of equality represented by the solid line. Bias between the methods is represented as the mean of the difference (+0.21), with the upper and lower 95% LoA plotted as +1.96 s (+0.50) and −1.96 s (−0.07), respectively. IP–MS/MS, immunoprecipitation–tandem mass spectrometry; NfL, neurofilament light chain; s, standard deviation; Simoa, single-molecule array; UCL, University College London.
Figure 4
Figure 4
Relative differences in NfL concentration between clinical groups are conserved when measured by Simoa or IP–MS/MS (UCL). The relative differences in CSF NfL concentration between clinical groups as measured by (A) Simoa were found to be conserved when compared to (B) measurement of the Coil 2B NfL316–323 (TLEIEACR) peptide by IP–MS/MS (UCL). Significant differences in NfL concentrations between clinical diagnostic groups were determined by Mann–Whitney U-tests. For both NfL assays, all neurodegenerative diseases except the multiple sclerosis group were found to be significantly different to healthy controls: dementia with Lewy bodies (P < 0.01), Alzheimer’s disease (P ≤ 0.01), semantic dementia (P = 0.001), corticobasal syndrome (P < 0.05), behavioural variant frontotemporal dementia (P < 0.05) and Huntington’s disease (P < 0.0005). Ad, Alzheimer’s disease; bvFTD, behavioural variant frontotemporal dementia; CBS, corticobasal syndrome; DLB, dementia with Lewy bodies; HD, Huntington’s disease; IQT, interquartile range; MS, multiple sclerosis; NfL, neurofilament light chain; SD, semantic dementia.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Abu-Rumeileh S, Abdelhak A, Foschi M, et al. The multifaceted role of neurofilament light chain protein in non-primary neurological diseases. Brain. 2023;146(2):421–437. - PMC - PubMed
    1. Fisse AL, Pitarokoili K, Leppert D, et al. Serum neurofilament light chain as outcome marker for intensive care unit patients. J Neurol. 2021;268(4):1323–1329. - PMC - PubMed
    1. Taccone FS, Nolan JP, Hoedemaekers CWE. Neurofilament to predict post-anoxic neurological outcome: Are we ready for the prime time? Intensive Care Med. 2021;47(1):77–79. - PubMed
    1. Masvekar RR, Kosa P, Jin K, et al. Prognostic value of serum/plasma neurofilament light chain for COVID-19-associated mortality. Ann Clin Transl Neurol. 2022;9(5):622–632. - PMC - PubMed
    1. Virhammar J, Naas A, Fallmar D, et al. Biomarkers for central nervous system injury in cerebrospinal fluid are elevated in COVID-19 and associated with neurological symptoms and disease severity. Eur J Neurol. 2021;28(10):3324–3331. - PMC - PubMed

LinkOut - more resources

-