TY - JOUR
T1 - A Novel Neurofilament Light Chain ELISA Validated in Patients with Alzheimer’s Disease, Frontotemporal Dementia, and Subjective Cognitive Decline, and the Evaluation of Candidate Proteins for Immunoassay Calibration
AU - Das, Shreyasee
AU - Dewit, Nele
AU - Jacobs, Dirk
AU - Pijnenburg, Yolande A. L.
AU - in ‘t Veld, Sjors G. J. G.
AU - Coppens, Salomé
AU - Quaglia, Milena
AU - Hirtz, Christophe
AU - Teunissen, Charlotte E.
AU - Vanmechelen, Eugeen
N1 - Funding Information: Conflicts of Interest: S.D. and D.J. are employees of ADx NeuroSciences, Gent, Belgium. S.D. is also enrolled as an external PhD candidate at the Amsterdam University Medical Centre, Amsterdam, The Netherlands. E.V.M. is the co-founder of ADx NeuroSciences. N.D. was an employee of ADx NeuroSciences at the time of drafting the manuscript and is currently employed at Medpace, Leu-ven, Belgium. C.E.T. has a collaboration contract with ADx NeuroSciences, Quanterix and Eli Lilly, performed contract research or received grants from AC-Immune, Axon Neurosciences, Bioconnect, Bioor-chestra, Brainstorm Therapeutics, Celgene, EIP Pharma, Eisai, Grifols, Novo Nordisk, Peo-pleBio, Roche, Toyama, Vivoryon. She serves on editorial boards of Medidact Neurologie/Springer, Alzheimer Research and Therapy, Neurology: Neuroimmunology & Neuroinflammation, and is editor of a Neuromethods book Springer. All of the other authors declare no competing interests. Funding Information: Funding: The research discussed herein has received funding from the European Union’s Horizon 2020 research and innovation program Marie Curie ITN ‘MIRIADE’ under the grant agreement number 860303. Part of the data used in this article has been collected as a part of the 18HLT09 Neu-roMET2 project, which received funding from the EMPIR program co-financed by the Participating States and from the European Union’s Horizon 2020 research and innovation program. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/7/1
Y1 - 2022/7/1
N2 - Neurofilament light chain (Nf-L) is a well-known biomarker for axonal damage; however, the corresponding circulating Nf-L analyte in cerebrospinal fluid (CSF) is poorly characterized. We therefore isolated new monoclonal antibodies against synthetic peptides, and these monoclonals were characterized for their specificity on brain-specific intermediate filament proteins. Two highly specific antibodies, ADx206 and ADx209, were analytically validated for CSF applications according to well-established criteria. Interestingly, using three different sources of purified Nf-L proteins, a significant impact on interpolated concentrations was observed. With a lower limit of analytical sensitivity of 100 pg/mL using bovine Nf-L as the calibrator, we were able to quantify the Nf-L analyte in each sample, and these Nf-L concentrations were highly correlated to the Uman diagnostics assay (Spearman rho = 0.97, p < 0.001). In the clinical diagnostic groups, the new Nf-L ELISA could discriminate patients with Alzheimer’s disease (AD, n = 20) from those with frontotemporal lobe dementia (FTD, n = 20) and control samples with subjective cognitive decline (SCD, n = 20). Hence-forth, this novel Nf-L ELISA with well-defined specificity and epitopes can be used to enhance our understanding of harmonizing the use of Nf-L as a clinically relevant marker for neurodegeneration in CSF.
AB - Neurofilament light chain (Nf-L) is a well-known biomarker for axonal damage; however, the corresponding circulating Nf-L analyte in cerebrospinal fluid (CSF) is poorly characterized. We therefore isolated new monoclonal antibodies against synthetic peptides, and these monoclonals were characterized for their specificity on brain-specific intermediate filament proteins. Two highly specific antibodies, ADx206 and ADx209, were analytically validated for CSF applications according to well-established criteria. Interestingly, using three different sources of purified Nf-L proteins, a significant impact on interpolated concentrations was observed. With a lower limit of analytical sensitivity of 100 pg/mL using bovine Nf-L as the calibrator, we were able to quantify the Nf-L analyte in each sample, and these Nf-L concentrations were highly correlated to the Uman diagnostics assay (Spearman rho = 0.97, p < 0.001). In the clinical diagnostic groups, the new Nf-L ELISA could discriminate patients with Alzheimer’s disease (AD, n = 20) from those with frontotemporal lobe dementia (FTD, n = 20) and control samples with subjective cognitive decline (SCD, n = 20). Hence-forth, this novel Nf-L ELISA with well-defined specificity and epitopes can be used to enhance our understanding of harmonizing the use of Nf-L as a clinically relevant marker for neurodegeneration in CSF.
KW - ELISA
KW - biomarker
KW - calibrator
KW - immunoassay
KW - neurofilament-light
UR - http://www.scopus.com/inward/record.url?scp=85133018038&partnerID=8YFLogxK
U2 - https://doi.org/10.3390/ijms23137221
DO - https://doi.org/10.3390/ijms23137221
M3 - Article
C2 - 35806226
SN - 1661-6596
VL - 23
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 13
M1 - 7221
ER -