Neurofilament Light Chain under the Lens of Structural Mass Spectrometry

Salomé Coppens, Dea Gogishvili, Valentina Faustinelli, Emanuele Scollo, Christopher Hopley, Sanne Abeln, Paul Dalby, Heidi Goenaga-Infante*, Luise Luckau, Jérôme Vialaret, Sylvain Lehmann, Christophe Hirtz, Eva Illes-Toth

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Neurofilament light chain (NfL) is an early nonspecific biomarker in neurodegenerative diseases and traumatic brain injury, indicating axonal damage. This work describes the detailed structural characterization of a selected primary calibrator with the potential to be used in future reference measurement procedure (RMP) development for the accurate quantification of NfL. As a part of the described workflow, the sequence, higher-order structure as well as solvent accessibility, and hydrogen-bonding profile were assessed under three different conditions in KPBS, artificial cerebrospinal fluid, and artificial cerebrospinal fluid in the presence of human serum albumin. The results revealed that NfL is a structurally heterogeneous protein, eliciting a large conformational flexibility. Its structural ensemble changed when it was diluted with an aqueous buffer versus a surrogate matrix, artificial cerebrospinal fluid (aCSF), and/or aCSF with human serum albumin. Various regions of protection and deprotection in the protein head, central helical, and tail domains that experienced altered solvent accessibility and conformational changes caused by different solvent conditions were identified. Moreover, interfacial residues, which may play a role in a potential direct interaction between NfL and human serum albumin, emerged from hydrogen-deuterium exchange mass spectrometry (HDX-MS). These data pinpointed distinct regions of the protein that may participate in such an interaction. Overall, critical quality attributes of a potential primary calibrator for NfL measurements are provided. These findings will ultimately inform ongoing biochemical and clinical assay development procedures and manufacturing practices, giving careful consideration during sample handling and method development.

Original languageEnglish
Pages (from-to)141-151
Number of pages11
JournalACS Chemical Neuroscience
Volume16
Issue number2
DOIs
Publication statusPublished - 15 Jan 2025

Bibliographical note

Publisher Copyright:
© 2025 The Authors. Published by American Chemical Society.

Keywords

  • human serum albumin
  • hydrogen−deuterium exchange
  • mass spectrometry
  • neurofilament light chain
  • primary calibrator
  • structural characterization

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