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A specific ELISA for measuring neurofilament heavy chain phosphoforms
A Petzold1, G Keir, A J E Green
1Department of Neuroimmunology, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK. a.petzold@ion.ucl.ac.uk
Journal of Immunological Methods
|September 6, 2003
Summary
A new enzyme-linked immunosorbent assay (ELISA) quantifies neurofilament (Nf) levels in bodily fluids. This tool helps detect axonal degeneration, showing higher NfH(SMI35) levels in neurological diseases like ALS.
Area of Science:
- Neuroscience
- Biochemistry
Background:
- Neurofilaments (Nf) are key components of the neuronal cytoskeleton.
- Measuring Nf levels in body fluids offers insights into axonal degeneration.
- Specific Nf phosphoforms are crucial biomarkers for neurological conditions.
Purpose of the Study:
- To develop and validate a novel sandwich ELISA for quantifying the NfH(SMI35) phosphoform.
- To establish the assay's sensitivity, precision, and stability in various biological samples.
- To determine reference values for NfH(SMI35) in cerebrospinal fluid (CSF) and assess its diagnostic utility.
Main Methods:
- Development of a sandwich ELISA for NfH(SMI35) quantification.
- Validation of the assay using CSF, brain tissue, and cell culture homogenates.
- Determination of assay sensitivity, recovery, precision, and sample stability.
- Establishment of upper reference limits using a large cohort of CSF samples.
Main Results:
- The NfH(SMI35) ELISA demonstrated high sensitivity (0.2 ng/ml) and good recovery.
- CSF NfH(SMI35) levels were stable under specific storage conditions and resistant to freeze-thaw cycles.
- Elevated NfH(SMI35) levels were observed in patients with amyotrophic lateral sclerosis, space-occupying lesions, disc prolapse, and subarachnoid hemorrhage.
- A proposed nomenclature facilitates comparison across different detection methods.
Conclusions:
- The developed NfH(SMI35) ELISA is a sensitive and reliable tool for quantifying axonal degeneration.
- CSF NfH(SMI35) serves as a valuable biomarker for diagnosing and monitoring specific neurological disorders.
- The assay's robustness and established reference values support its clinical application.