Measuring neurofilament light in human plasma and cerebrospinal fluid: a comparison of five analytical immunoassays

Udit Sheth^{1

2}, Rebecca Harrison³, Kyle Ferber⁴, Erin G Rosenbaugh⁵, Amanda Bevis⁵, Rohini Khillan⁵, Michael Benatar⁶, Nicole L Bjorklund⁷, Elena Di Daniel³, Glenn A Harris⁸, Olga I Kahn⁹, Yongge Liu¹⁰, Henrik Zetterberg^{11

12

13

14

15

16}, Laura L Mitic¹⁷, Danielle Graham¹⁸, Tania F Gendron^{1

2}

Affiliations

¹ Department of Neuroscience, Mayo Clinic, Jacksonville, FL, USA.
² Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Jacksonville, FL, USA.
³ Astex Pharmaceuticals, Cambridge, UK.
⁴ Department of Biostatistics, Biogen, Cambridge, MA, USA.
⁵ Foundation at the National Institute of Health Inc., North Bethesda, MD, USA.
⁶ Department of Neurology and ALS Center, University of Miami Miller School of Medicine, Miami, FL, USA.
⁷ Association for Frontotemporal Degeneration, King of Prussia, PA, USA.
⁸ Rainwater Charitable Foundation, Fort Worth, TX, USA.
⁹ Alector, Inc., South San Francisco, CA, USA.
¹⁰ Otsuka Pharmaceutical Development & Commercialization, Inc, Rockville, MD, USA.
¹¹ Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, The Sahlgrenska Academy at the University of Gothenburg, Mölndal, Sweden.
¹² Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, Mölndal, Sweden.
¹³ Department of Neurodegenerative Disease, UCL Institute of Neurology, London, UK.
¹⁴ UK Dementia Research Institute at UCL, London, UK.
¹⁵ Hong Kong Center for Neurodegenerative Diseases, InnoHK, Hong Kong, China.
¹⁶ Wisconsin Alzheimer's Disease Research Center, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, USA.
¹⁷ The Bluefield Project to Cure Frontotemporal Dementia and the University of California, San Francisco, CA, USA.
¹⁸ Department of Biomarkers and Systems Biology, Biogen, Cambridge, MA, USA.

PMID: 40958756
DOI: 10.1515/cclm-2025-0610

Measuring neurofilament light in human plasma and cerebrospinal fluid: a comparison of five analytical immunoassays

Udit Sheth et al. Clin Chem Lab Med. 2025.

. 2025 Sep 8.

doi: 10.1515/cclm-2025-0610. Online ahead of print.

Authors

Affiliations

¹ Department of Neuroscience, Mayo Clinic, Jacksonville, FL, USA.
² Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Jacksonville, FL, USA.
³ Astex Pharmaceuticals, Cambridge, UK.
⁴ Department of Biostatistics, Biogen, Cambridge, MA, USA.
⁵ Foundation at the National Institute of Health Inc., North Bethesda, MD, USA.
⁶ Department of Neurology and ALS Center, University of Miami Miller School of Medicine, Miami, FL, USA.
⁷ Association for Frontotemporal Degeneration, King of Prussia, PA, USA.
⁸ Rainwater Charitable Foundation, Fort Worth, TX, USA.
⁹ Alector, Inc., South San Francisco, CA, USA.
¹⁰ Otsuka Pharmaceutical Development & Commercialization, Inc, Rockville, MD, USA.
¹¹ Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, The Sahlgrenska Academy at the University of Gothenburg, Mölndal, Sweden.
¹² Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, Mölndal, Sweden.
¹³ Department of Neurodegenerative Disease, UCL Institute of Neurology, London, UK.
¹⁴ UK Dementia Research Institute at UCL, London, UK.
¹⁵ Hong Kong Center for Neurodegenerative Diseases, InnoHK, Hong Kong, China.
¹⁶ Wisconsin Alzheimer's Disease Research Center, University of Wisconsin School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, USA.
¹⁷ The Bluefield Project to Cure Frontotemporal Dementia and the University of California, San Francisco, CA, USA.
¹⁸ Department of Biomarkers and Systems Biology, Biogen, Cambridge, MA, USA.

PMID: 40958756
DOI: 10.1515/cclm-2025-0610

Abstract

Objectives: Neurofilament light (NfL) is an established biofluid marker of neuroaxonal injury for neurological diseases. Several high-throughput and sensitive immunoassays have been developed to quantify NfL in blood and cerebrospinal fluid (CSF), facilitating the use of NfL as a biomarker in research and clinical practice. However, because of the lack of rigorous comparisons of assays, it has been difficult to determine whether data are comparable and whether assay performance differs. Here, we compared the performance of five NfL immunoassays.

Methods: To assess the five NfL immunoassays (Fujirebio, ProteinSimple, Quanterix, Roche and Siemens), we used pooled plasma or pooled CSF, as well as unique samples from 20 healthy controls and 20 individuals with El Escorial defined probable or definite amyotrophic lateral sclerosis (ALS), to evaluate precision, parallelism and/or bias. We also examined correlations between plasma and CSF NfL concentrations within and across assays and evaluated their ability to differentiate healthy controls from individuals with ALS.

Results: Four of the five assays demonstrated exemplary performance based on our analyses of precision and parallelism. Across the five assays, NfL concentrations were lower in plasma than in CSF, although they displayed a high degree of correlation. We noted bias across assays; plasma NfL concentrations were lowest for the Roche assay and highest for the ProteinSimple assay. In addition, all assays reliably distinguished healthy controls from individuals with ALS using plasma or CSF NfL.

Conclusions: Four NfL assays demonstrated similar analytic performance. Alongside performance, other factors such as costs, accessibility, usability, footprint, and intended use, should be considered.

Keywords: amyotrophic lateral sclerosis; biomarker; cerebrospinal fluid; immunoassays; neurofilament light chain; plasma.

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Update of

Measuring neurofilament light in human plasma and cerebrospinal fluid: a comparison of five analytical immunoassays.
Sheth U, Harrison R, Ferber K, Rosenbaugh EG, Bevis A, Khillan R, Benatar M, Bjorklund NL, Di Daniel E, Harris GA, Kahn OI, Liu Y, Zetterberg H, Mitic LL, Graham D, Gendron TF. Sheth U, et al. bioRxiv [Preprint]. 2025 May 9:2025.05.05.652212. doi: 10.1101/2025.05.05.652212. bioRxiv. 2025. Update in: Clin Chem Lab Med. 2025 Sep 08. doi: 10.1515/cclm-2025-0610. PMID: 40654957 Free PMC article. Updated. Preprint.

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Measuring neurofilament light in human plasma and cerebrospinal fluid: a comparison of five analytical immunoassays

Affiliations

Measuring neurofilament light in human plasma and cerebrospinal fluid: a comparison of five analytical immunoassays

Authors

Affiliations

Abstract

Update of

References

LinkOut - more resources

Full Text Sources

Miscellaneous