A new enzyme-linked immunosorbent assay for neurofilament light in cerebrospinal fluid: Analytical validation and clinical evaluation

Lorenzo Gaetani, Kina Höglund, Lucilla Parnetti, Fani Pujol-Calderon, Bruno Becker, Paolo Eusebi, Paola Sarchielli, Paolo Calabresi, Massimiliano Di Filippo, Henrik Zetterberg, Kaj Blennow

Research output: Contribution to journalArticle

Abstract

Background: Cerebrospinal fluid (CSF) neurofilament light (NfL) is a reliable marker of neuro-axonal damage in different neurological disorders that is related to disease severity. To date, all recent studies performed in human CSF have used the same enzyme-linked immunosorbent assay (ELISA). To confirm the large body of evidence for NfL, we developed a new ELISA method and here we present the performance characteristics of this new ELISA for CSF NfL in different neurological disorders. Methods: We produced two monoclonal antibodies (NfL21 and NfL23) directed against the NfL core domain, and developed a novel sandwich ELISA method that we evaluated in patients with: 1) inflammatory demyelinating diseases (IDD; n = 97), including multiple sclerosis (MS; n = 59), clinically isolated syndrome (CIS; n = 32), and radiologically isolated syndrome (RIS; n = 6); 2) Alzheimer's disease (AD; n = 72), including mild cognitive impairment due to AD (MCI-AD, n = 36) and probable AD dementia (AD-dem; n = 36); 3) Parkinson's disease (PD; n = 30); and 4) other neurological noninflammatory and non-neurodegenerative diseases (OND; n = 30). Results: Our new NfL ELISA showed a good analytical performance (inter-plate coefficient of variation (CV) < 13%), with no cross-reactivity with neurofilament medium and heavy (NfM and NfH). With respect to the other available ELISAs, CSF NfL showed the same range of values with a strong correlation (r = 0.9984, p < 0.001) between the two methods. CSF NfL levels were significantly higher in MCI-AD/AD-dem and IDD patients as compared with both PD and OND patients. The highest discriminative power was obtained between IDD and OND patients (area under the curve (AUC) 0.87, 95% confidence interval (CI) 0.80-0.95). Within the IDD group, CSF NfL positively correlated with several clinical and radiological disease severity parameters. Conclusions: These results show a good analytical performance of the new ELISA for quantification of NfL concentrations in the CSF. CSF NfL is confirmed to be a reliable marker in AD and MS, and a disease-severity marker in MS patients.

Original languageEnglish
Article number8
JournalAlzheimer's Research and Therapy
Volume10
Issue number1
DOIs
Publication statusPublished - Jan 23 2018

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Intermediate Filaments
Cerebrospinal Fluid
Enzyme-Linked Immunosorbent Assay
Light
Nervous System Diseases
Demyelinating Diseases
Multiple Sclerosis
Area Under Curve
Dementia
Monoclonal Antibodies
Confidence Intervals

Keywords

  • Alzheimer's disease
  • Cerebrospinal fluid
  • Clinically isolated syndrome
  • ELISA
  • Mild cognitive impairment
  • Multiple sclerosis
  • Neurofilament light
  • Parkinson's disease

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology
  • Cognitive Neuroscience

Cite this

A new enzyme-linked immunosorbent assay for neurofilament light in cerebrospinal fluid : Analytical validation and clinical evaluation. / Gaetani, Lorenzo; Höglund, Kina; Parnetti, Lucilla; Pujol-Calderon, Fani; Becker, Bruno; Eusebi, Paolo; Sarchielli, Paola; Calabresi, Paolo; Di Filippo, Massimiliano; Zetterberg, Henrik; Blennow, Kaj.

In: Alzheimer's Research and Therapy, Vol. 10, No. 1, 8, 23.01.2018.

Research output: Contribution to journalArticle

Gaetani, L, Höglund, K, Parnetti, L, Pujol-Calderon, F, Becker, B, Eusebi, P, Sarchielli, P, Calabresi, P, Di Filippo, M, Zetterberg, H & Blennow, K 2018, 'A new enzyme-linked immunosorbent assay for neurofilament light in cerebrospinal fluid: Analytical validation and clinical evaluation', Alzheimer's Research and Therapy, vol. 10, no. 1, 8. https://doi.org/10.1186/s13195-018-0339-1
Gaetani, Lorenzo ; Höglund, Kina ; Parnetti, Lucilla ; Pujol-Calderon, Fani ; Becker, Bruno ; Eusebi, Paolo ; Sarchielli, Paola ; Calabresi, Paolo ; Di Filippo, Massimiliano ; Zetterberg, Henrik ; Blennow, Kaj. / A new enzyme-linked immunosorbent assay for neurofilament light in cerebrospinal fluid : Analytical validation and clinical evaluation. In: Alzheimer's Research and Therapy. 2018 ; Vol. 10, No. 1.
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T1 - A new enzyme-linked immunosorbent assay for neurofilament light in cerebrospinal fluid

T2 - Analytical validation and clinical evaluation

AU - Gaetani, Lorenzo

AU - Höglund, Kina

AU - Parnetti, Lucilla

AU - Pujol-Calderon, Fani

AU - Becker, Bruno

AU - Eusebi, Paolo

AU - Sarchielli, Paola

AU - Calabresi, Paolo

AU - Di Filippo, Massimiliano

AU - Zetterberg, Henrik

AU - Blennow, Kaj

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N2 - Background: Cerebrospinal fluid (CSF) neurofilament light (NfL) is a reliable marker of neuro-axonal damage in different neurological disorders that is related to disease severity. To date, all recent studies performed in human CSF have used the same enzyme-linked immunosorbent assay (ELISA). To confirm the large body of evidence for NfL, we developed a new ELISA method and here we present the performance characteristics of this new ELISA for CSF NfL in different neurological disorders. Methods: We produced two monoclonal antibodies (NfL21 and NfL23) directed against the NfL core domain, and developed a novel sandwich ELISA method that we evaluated in patients with: 1) inflammatory demyelinating diseases (IDD; n = 97), including multiple sclerosis (MS; n = 59), clinically isolated syndrome (CIS; n = 32), and radiologically isolated syndrome (RIS; n = 6); 2) Alzheimer's disease (AD; n = 72), including mild cognitive impairment due to AD (MCI-AD, n = 36) and probable AD dementia (AD-dem; n = 36); 3) Parkinson's disease (PD; n = 30); and 4) other neurological noninflammatory and non-neurodegenerative diseases (OND; n = 30). Results: Our new NfL ELISA showed a good analytical performance (inter-plate coefficient of variation (CV) < 13%), with no cross-reactivity with neurofilament medium and heavy (NfM and NfH). With respect to the other available ELISAs, CSF NfL showed the same range of values with a strong correlation (r = 0.9984, p < 0.001) between the two methods. CSF NfL levels were significantly higher in MCI-AD/AD-dem and IDD patients as compared with both PD and OND patients. The highest discriminative power was obtained between IDD and OND patients (area under the curve (AUC) 0.87, 95% confidence interval (CI) 0.80-0.95). Within the IDD group, CSF NfL positively correlated with several clinical and radiological disease severity parameters. Conclusions: These results show a good analytical performance of the new ELISA for quantification of NfL concentrations in the CSF. CSF NfL is confirmed to be a reliable marker in AD and MS, and a disease-severity marker in MS patients.

AB - Background: Cerebrospinal fluid (CSF) neurofilament light (NfL) is a reliable marker of neuro-axonal damage in different neurological disorders that is related to disease severity. To date, all recent studies performed in human CSF have used the same enzyme-linked immunosorbent assay (ELISA). To confirm the large body of evidence for NfL, we developed a new ELISA method and here we present the performance characteristics of this new ELISA for CSF NfL in different neurological disorders. Methods: We produced two monoclonal antibodies (NfL21 and NfL23) directed against the NfL core domain, and developed a novel sandwich ELISA method that we evaluated in patients with: 1) inflammatory demyelinating diseases (IDD; n = 97), including multiple sclerosis (MS; n = 59), clinically isolated syndrome (CIS; n = 32), and radiologically isolated syndrome (RIS; n = 6); 2) Alzheimer's disease (AD; n = 72), including mild cognitive impairment due to AD (MCI-AD, n = 36) and probable AD dementia (AD-dem; n = 36); 3) Parkinson's disease (PD; n = 30); and 4) other neurological noninflammatory and non-neurodegenerative diseases (OND; n = 30). Results: Our new NfL ELISA showed a good analytical performance (inter-plate coefficient of variation (CV) < 13%), with no cross-reactivity with neurofilament medium and heavy (NfM and NfH). With respect to the other available ELISAs, CSF NfL showed the same range of values with a strong correlation (r = 0.9984, p < 0.001) between the two methods. CSF NfL levels were significantly higher in MCI-AD/AD-dem and IDD patients as compared with both PD and OND patients. The highest discriminative power was obtained between IDD and OND patients (area under the curve (AUC) 0.87, 95% confidence interval (CI) 0.80-0.95). Within the IDD group, CSF NfL positively correlated with several clinical and radiological disease severity parameters. Conclusions: These results show a good analytical performance of the new ELISA for quantification of NfL concentrations in the CSF. CSF NfL is confirmed to be a reliable marker in AD and MS, and a disease-severity marker in MS patients.

KW - Alzheimer's disease

KW - Cerebrospinal fluid

KW - Clinically isolated syndrome

KW - ELISA

KW - Mild cognitive impairment

KW - Multiple sclerosis

KW - Neurofilament light

KW - Parkinson's disease

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