Development and multi-center validation of a fully automated digital immunoassay for neurofilament light chain: toward a clinical blood test for neuronal injury

Author:

Wilson David1,Chan Dandan1,Chang Lei1,Mathis Robert1,Verberk Inge2,Montalban Xavier3,Comabella Manuel3,Fissolo Nicolas3,Bielekova Bibi4,Masvekar Ruturaj4,Chitnis Tanuja5,Ziemssen Tjalf6,Akgün Katja6,Blennow Kaj7,Zetterberg Henrik7891011,Brück Wolfgang12,Giovannoni Gavin13,Gnanapavan Sharmilee13,Bittner Stefan14,Zipp Frauke14,Comi Giancarlo15,Furlan Roberto15,Lehmann Sylvain16,Thebault Simon17,Freedman Mark17,Bar-Or Amit18,Kramer Marty19,Otto Markus20,Halbgebauer Steffen2021,Hrusovsky Kevin1,Plavina Tatiana1,Khalil Michael22,Piehl Fredrik23,Wiendl Heinz24,Kappos Ludwig25,Maceski Aleksandra26,Willemse Eline26,Leppert David26,Teunissen Charlotte2,Kuhle Jens26

Affiliation:

1. Quanterix Corp , Billerica , MA , USA

2. Neurochemistry Laboratory, Department of Laboratory Medicine, Amsterdam Neuroscience, Amsterdam , University Medical Centers , Amsterdam , The Netherlands

3. Laboratori de Neuroinmunologia Clinica Centre d’Esclerosi Múltiple de Catalunya (Cemcat) Vall d’Hebron Institut de Recerca , Barcelona , Spain

4. National Institute of Allergy and Infectious Diseases, National Institutes of Health , Bethesda , MD , USA

5. Brigham and Women’s Hospital, Harvard Medical School , Boston , MA , USA

6. MS Center Dresden, Center of Clinical Neuroscience, Department of Neurology , Dresden University of Technology , Dresden , Germany

7. Department of Psychiatry and Neurochemistry , Institute of Neuroscience and Physiology, The Sahlgrenska Academy at the University of Gothenburg , Mölndal , Sweden

8. Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital , Mölndal , Sweden

9. Department of Neurodegenerative Disease , UCL Institute of Neurology, Queen Square , London , UK

10. UK Dementia Research Institute at UCL , London , UK

11. Hong Kong Center for Neurodegenerative Diseases, Clear Water Bay , Hong Kong , China

12. Institute for Neuropathology at the University Medical Center , Göttingen , Germany

13. Department of Neurology , Barts Health NHS Trust, The Royal London Hospital, E1 1FR , London , UK

14. University Medical Center Mainz , Department of Neurology , Mainz , Germany

15. Institute of Experimental Neurology, Division of Neuroscience, University Vita e Salute San Raffaele and IRCCS San Raffaele Hospital , Milan , Italy

16. Hôpital St Eloi , Montpellier , France

17. University of Ottawa , Department of Medicine, The Ottawa Hospital Research Institute , Ottawa , Canada

18. Perelman School of Medicine, University of Pennsylvania , Philadelphia , PA , USA

19. Sanofi , Framingham , MA , USA

20. Department of Neurology , Ulm University Hospital , Ulm , Germany

21. German Center for Neurodegenerative Diseases (DZNE e.V.) , Ulm , Germany

22. Department of Neurology , Medical University of Graz , Graz , Austria

23. Department of Clinical Neuroscience , Karolinska Institutet , Stockholm , Sweden

24. Department of Neurology , University of Münster , Münster , Germany

25. Neurologic Clinic and Policlinic, MS Center and Research Center for Clinical Neuroimmunology and Neuroscience Basel, Departments of Biomedicine and Clinical Research , University Hospital Basel, University of Basel , Basel , Switzerland

26. Multiple Sclerosis Centre and Research Center for Clinical Neuroimmunology and Neuroscience Basel (RC2NB), Departments of Head, Spine and Neuromedicine, Biomedicine and Clinical Research , University Hospital Basel and University of Basel , Basel , Switzerland

Abstract

Abstract Objectives Neurofilament light chain (NfL) has emerged as a promising biomarker for detecting and monitoring axonal injury. Until recently, NfL could only be reliably measured in cerebrospinal fluid, but digital single molecule array (Simoa) technology has enabled its precise measurement in blood samples where it is typically 50–100 times less abundant. We report development and multi-center validation of a novel fully automated digital immunoassay for NfL in serum for informing axonal injury status. Methods A 45-min immunoassay for serum NfL was developed for use on an automated digital analyzer based on Simoa technology. The analytical performance (sensitivity, precision, reproducibility, linearity, sample type) was characterized and then cross validated across 17 laboratories in 10 countries. Analytical performance for clinical NfL measurement was examined in individual patients with relapsing remitting multiple sclerosis (RRMS) after 3 months of disease modifying treatment (DMT) with fingolimod. Results The assay exhibited a lower limit of detection (LLoD) of 0.05 ng/L, a lower limit of quantification (LLoQ) of 0.8 ng/L, and between-laboratory imprecision <10 % across 17 validation sites. All tested samples had measurable NfL concentrations well above the LLoQ. In matched pre–post treatment samples, decreases in NfL were observed in 26/29 RRMS patients three months after DMT start, with significant decreases detected in a majority of patients. Conclusions The sensitivity characteristics and reproducible performance across laboratories combined with full automation make this assay suitable for clinical use for NfL assessment, monitoring in individual patients, and cross-comparisons of results across multiple sites.

Publisher

Walter de Gruyter GmbH

Subject

Biochemistry (medical),Clinical Biochemistry,General Medicine

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