Quantitation of neurofilament light chain protein in serum and cerebrospinal fluid from patients with multiple sclerosis using the MSD R-PLEX NfL assay

Antigona Ulndreaj; Dorsa Sohaei; Simon Thebault; Oscar D. Pons-Belda; Amaia Fernandez-Uriarte; Christopher Campbell; David Cheo; Martin Stengelin; George Sigal; Mark S. Freedman; Isobel A. Scarisbrick; Ioannis Prassas; Eleftherios P. Diamandis

doi:10.1515/dx-2022-0125

Article

Quantitation of neurofilament light chain protein in serum and cerebrospinal fluid from patients with multiple sclerosis using the MSD R-PLEX NfL assay

Antigona Ulndreaj , Dorsa Sohaei , Simon Thebault , Oscar D. Pons-Belda , Amaia Fernandez-Uriarte , Christopher Campbell , David Cheo , Martin Stengelin , George Sigal , Mark S. Freedman , Isobel A. Scarisbrick , Ioannis Prassas and Eleftherios P. Diamandis

Published/Copyright: February 15, 2023

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From the journal Diagnosis Volume 10 Issue 3

Abstract

Objectives

Neurofilament light (NfL) chain is a marker of neuroaxonal damage in various neurological diseases. Here we quantitated NfL levels in the cerebrospinal fluid (CSF) and serum from patients with multiple sclerosis (MS) and controls, using the R-PLEX NfL assay, which employs advanced Meso Scale Discovery^® (MSD) electrochemiluminescence (ECL)-based detection technology.

Methods

NfL was quantitated in samples from 116 individuals from two sites (Ottawa Hospital Research Institute and Mayo Clinic), consisting of patients with MS (n=71) and age- and sex-matched inflammatory neurological controls (n=13) and non-inflammatory controls (n=32). Correlation of NfL levels between CSF and serum was assessed in paired samples in a subset of MS patients and controls (n=61). Additionally, we assessed the correlation between NfL levels obtained with MSD’s R-PLEX^® and Quanterix’s single molecule array (Simoa^®) assays in CSF and serum (n=32).

Results

Using the R-PLEX, NfL was quantitated in 99% of the samples tested, and showed a broad range in the CSF (82–500,000 ng/L) and serum (8.84–2,014 ng/L). Nf-L levels in both biofluids correlated strongly (r=0.81, p<0.0001). Lastly, Nf-L measured by MSD’s R-PLEX and Quanterix’s Simoa assays were highly correlated for both biofluids (CSF: r=0.94, p<0.0001; serum: r=0.95, p<0.0001).

Conclusions

We show that MSD’s R-PLEX NfL assay can reliably quantitate levels of NfL in the CSF and serum from patients with MS and controls, where levels correlate strongly with Simoa.

Keywords: cerebrospinal fluid; electrochemiluminescence; multiple sclerosis; neurofilament light; R-PLEX; serum; Simoa

Corresponding authors: Dr. Ioannis Prassas, PhD, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Joseph & Wolf Lebovic Ctr., 60 Murray St [Box 32]; Flr 6 – Rm L6-201, Toronto, ON M5T 3L9, Canada; and Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON, Canada, Phone: 416 586 8443, Fax: 416 619 5521, E-mail: yprassas@gmail.com; and Dr. Eleftherios P. Diamandis, PhD, MD, Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Joseph & Wolf Lebovic Ctr., 60 Murray St [Box 32]; Floor 6 – Rm L6-201, Toronto, ON M5T 3L9, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada; and Department of Clinical Biochemistry, University Health Network, Toronto, ON, Canada, Phone: 416 586 8443, Fax: 416 619 5521, E-mail: Eleftherios.Diamandis@sinaihealth.ca

Funding source: National Institute of Allergy and Infectious Diseases

Award Identifier / Grant number: U24AI118660

Funding source: Ottawa Hospital Innovation Grant

Funding source: University of Toronto

Award Identifier / Grant number: Unassigned

Funding source: Canada First Research Excellence Fund

Award Identifier / Grant number: Unassigned

Acknowledgments

Dr. Ulndreaj is supported by a post-doctoral fellowship from the University of Toronto’s Medicine by Design initiative, which receives funding from the Canada First Research Excellence Fund (CFREF).

Research funding: The development of the MSD assays was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under Award Number U24AI118660. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Funding for the Quanterix assays was provided by the Ottawa Hospital Innovation Grant.
Author contribution: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Some authors (CC, MS, GS, DC) are employees of MSD. Dr. Eleftherios P. Diamandis declares that he holds an advisory role with Abbott Diagnostics and a consultant role with Imaware Diagnostics. All other authors have nothing to declare.
Informed consent: Informed consent was obtained from all individuals included in this study.
Ethical approval: Research involving human subjects complied with all relevant national regulations, institutional policies and is in accordance with the tenets of the Helsinki Declaration (as revised in 2013) and has been approved by the authors’ Institutional Review Boards (Mount Sinai Hospital, The Ottawa Hospital Research Institute, and Mayo Clinic).

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Received: 2022-11-19

Revised: 2023-02-01

Accepted: 2023-02-05

Published Online: 2023-02-15

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Articles in the same Issue

https://doi.org/10.1515/dx-2022-0125

Keywords for this article

cerebrospinal fluid; electrochemiluminescence; multiple sclerosis; neurofilament light; R-PLEX; serum; Simoa