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Neurofilament Light Chain under the Lens of Structural Mass Spectrometry
Salomé Coppens1,2, Dea Gogishvili3,4, Valentina Faustinelli1,5
1National Measurement Laboratory, LGC, Queens Road, TW11 0LY Teddington, U.K.
ACS Chemical Neuroscience
|January 2, 2025
Summary
Neurofilament light chain (NfL) is a key biomarker for axonal damage. This study characterizes NfL structure, revealing its conformational flexibility and how it changes in different biological fluids, crucial for accurate NfL measurement development.
Area of Science:
- Biochemistry
- Structural Biology
- Biomarker Discovery
Background:
- Neurofilament light chain (NfL) serves as a nonspecific biomarker for neurodegenerative diseases and traumatic brain injury, indicating axonal damage.
- Accurate quantification of NfL is essential for reliable clinical diagnostics and research.
Purpose of the Study:
- To perform detailed structural characterization of a primary calibrator for Neurofilament light chain (NfL).
- To assess the impact of different solvent conditions on NfL structure and conformational flexibility.
- To identify critical quality attributes for NfL calibrator development.
Main Methods:
- Hydrogen-deuterium exchange mass spectrometry (HDX-MS) was employed to analyze NfL structure.
- Structural characterization included assessment of sequence, higher-order structure, solvent accessibility, and hydrogen-bonding profiles.
- Experiments were conducted in KPBS, artificial cerebrospinal fluid (aCSF), and aCSF with human serum albumin.
Main Results:
- Neurofilament light chain (NfL) exhibits significant structural heterogeneity and conformational flexibility.
- NfL's structural ensemble is sensitive to dilution in different matrices, including aqueous buffers, aCSF, and aCSF with human serum albumin.
- Specific regions of NfL, including head, helical, and tail domains, showed altered solvent accessibility and conformation based on solvent conditions.
- HDX-MS identified interfacial residues potentially involved in NfL-human serum albumin interactions.
Conclusions:
- The study provides critical quality attributes for a potential primary NfL calibrator.
- Understanding NfL's structural behavior in various conditions is vital for accurate quantification and assay development.
- Findings will guide biochemical and clinical assay development, sample handling, and manufacturing practices for NfL measurements.
Keywords:
human serum albuminhydrogen−deuterium exchangemass spectrometryneurofilament light chainprimary calibratorstructural characterizationMore Related Videos
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