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Peer-reviewedCC BY 4.0Scientific Reports

Fresh-to-frozen comparability of serum neurofilament light chain across four immunoassays

  • Pavlina Kusnierova
  • Krystof Damian Svub
  • David Zeman
  • Michaela Sobkova
  • Sona Maresova
  • Karin Vilimova
  • Hana Slepcanova
  • David Stejskal
  • Kamila Zondra Revendova
Open at sourcePDFDOICite

Abstract

Serum neurofilament light chain (sNfL) is an increasingly used biomarker of neuroaxonal injury in multiple sclerosis (MS), but differences between immunoassays and sample handling may limit comparability of results. We compared paired fresh and frozen serum samples from 20 patients with MS using four immunoassays: Simoa, Elecsys, Atellica, and ELISA. Matched aliquots were stored at − 70 °C for 10 days and subjected to a single freeze–thaw cycle. Fresh-to-frozen differences varied significantly among platforms (Friedman χ² = 24.06, p < 0.001; Kendall’s W = 0.401). Median concentrations decreased after freezing with Simoa (–12.8%), Elecsys (–5.4%), and ELISA (–9.7%), whereas Atellica showed minimal change (+ 0.9%). Between-assay correlations were moderate to strong in fresh (Spearman ρ = 0.663–0.877) and frozen samples (ρ = 0.677–0.931), despite substantial differences in absolute concentrations. Assay-specific Simoa and Elecsys Z-scores were strongly correlated in fresh (ρ = 0.886) and frozen samples (ρ = 0.923; both p < 0.001), but systematic differences remained. These findings indicate assay-dependent fresh-to-frozen differences and support maintaining consistent analytical immunoassays and sample-processing conditions for longitudinal sNfL monitoring.

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Cite

BibTeXRIS
BibTeX
@article{kusnierova2026freshtofrozen,
  title = {{Fresh-to-frozen comparability of serum neurofilament light chain across four immunoassays}},
  author = {Pavlina Kusnierova and Krystof Damian Svub and David Zeman and Michaela Sobkova and Sona Maresova and Karin Vilimova and Hana Slepcanova and David Stejskal and Kamila Zondra Revendova},
  journal = {Scientific Reports},
  year = {2026},
  month = oct,
  doi = {10.1038/s41598-026-77730-z},
  url = {https://www.nature.com/articles/s41598-026-77730-z},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Fresh-to-frozen comparability of serum neurofilament light chain across four immunoassays
AU  - Pavlina Kusnierova
AU  - Krystof Damian Svub
AU  - David Zeman
AU  - Michaela Sobkova
AU  - Sona Maresova
AU  - Karin Vilimova
AU  - Hana Slepcanova
AU  - David Stejskal
AU  - Kamila Zondra Revendova
PY  - 2026
DA  - 2026/10/23
JO  - Scientific Reports
DO  - 10.1038/s41598-026-77730-z
UR  - https://www.nature.com/articles/s41598-026-77730-z
AB  - Serum neurofilament light chain (sNfL) is an increasingly used biomarker of neuroaxonal injury in multiple sclerosis (MS), but differences between immunoassays and sample handling may limit comparability of results. We compared paired fresh and frozen serum samples from 20 patients with MS using four immunoassays: Simoa, Elecsys, Atellica, and ELISA. Matched aliquots were stored at − 70 °C for 10 days and subjected to a single freeze–thaw cycle. Fresh-to-frozen differences varied significantly among platforms (Friedman χ² = 24.06, p < 0.001; Kendall’s W = 0.401). Median concentrations decreased after freezing with Simoa (–12.8%), Elecsys (–5.4%), and ELISA (–9.7%), whereas Atellica showed minimal change (+ 0.9%). Between-assay correlations were moderate to strong in fresh (Spearman ρ = 0.663–0.877) and frozen samples (ρ = 0.677–0.931), despite substantial differences in absolute concentrations. Assay-specific Simoa and Elecsys Z-scores were strongly correlated in fresh (ρ = 0.886) and frozen samples (ρ = 0.923; both p < 0.001), but systematic differences remained. These findings indicate assay-dependent fresh-to-frozen differences and support maintaining consistent analytical immunoassays and sample-processing conditions for longitudinal sNfL monitoring.
SN  - 2045-2322
ER  -