Seven day pre-analytical stability of serum and plasma neurofilament light chain
Open accessAbstract
Neurofilament light chain (NfL) has emerged as a biomarker of neuroaxonal damage in several neurologic conditions. With increasing availability of fourth-generation immunoassays detecting NfL in blood, aspects of pre-analytical stability of this biomarker remain unanswered. This study investigated NfL concentrations in serum and plasma samples of 32 patients with neurological diagnoses using state of the art Simoa technology. We tested the effect of delayed freezing of up to 7 days and statistically determined stability and validity of measured concentrations. We found concentrations of NfL in serum and plasma to remain stable at room temperature when processing of samples is delayed up to 7 days (serum: mean absolute difference 0.9 pg/mL, intraindividual variation 1.2%; plasma: mean absolute difference 0.5 pg/mL, intraindividual variation 1.3%). Consistency of these results was nearly perfect for serum and excellent for plasma (intraclass correlation coefficients 0.99 and 0.94, respectively). In conclusion, the soluble serum and plasma NfL concentration remains stable when unprocessed blood samples are stored up to 7 days at room temperature. This information is essential for ensuring reliable study protocols, for example, when shipment of fresh samples is needed.
Verification finished with gaps
Some checks didn’t finish. The results below cover the steps that did.
- Registry recordVerified
- Full textRead the full text
- Reference list15 references found
- Reference check15 of 15 references checked
- StatisticsRecomputed
- Claims30 claims anchored to a quote
- Cited sourcesChecked
- Self-checkDidn’t finish — the rest of the report still applies.
- VerificationFindings verified
AI snapshot
Written from the paper’s full text.
TL;DR
Serum and plasma neurofilament light chain (NfL) concentrations remained stable when unprocessed blood samples were stored at room temperature for up to 7 days before processing. The authors conclude that delayed processing is acceptable for NfL research protocols, with serum showing slightly better reliability than plasma.
Key findings
- For samples stored 7 days at room temperature, mean absolute difference versus immediate freezing was 0.9 pg/mL for serum NfL and 0.5 pg/mL for plasma NfL; intraindividual variation was 9.0% for serum and 6.5% for plasma.
- Reliability at 7 days at room temperature was near-perfect for serum (ICC = 0.99) and excellent for plasma (ICC = 0.94).
- Inter-assay variability was low: serum NfL iSD 0.6 pg/mL, iCOV 4.8%, ICC 1.0; plasma NfL iSD 1.6 pg/mL, iCOV 12.7%, ICC 0.99.
- Mean baseline concentrations were 14.5 pg/mL for serum NfL and 12.5 pg/mL for plasma NfL in 32 patients with neurological diagnoses.
Methods
- Serum and plasma samples from 32 patients with neurological diagnoses (multiple sclerosis n = 20, Parkinson’s disease n = 6, Alzheimer disease n = 4, primary headache n = 2) were collected in Greiner Bio-One Vacuette serum and EDTA tubes.
- Each participant provided 4 serum and 4 plasma samples stored before processing under four conditions: immediate, 3 days at 4–8 °C, 3 days at room temperature, and 7 days at room temperature.
- NfL was measured with Simoa Nf-light kits on a Simoa SR-X Analyzer; analyses included repeated-measures ANOVA, Bland–Altman plots, intraindividual SD/COV, and intraclass correlation coefficients. A separate validity cohort of 4 patients had paired serum/plasma samples measured on four assay plates for inter-assay reliability.
Limitations
- Stated: results may not generalize to other blood collection systems, since only Greiner Bio-One Vacuette tubes were used.
- Stated: validity depends on meticulous quality control in a specialized biomarker laboratory; real-world labs may have protocol deviations, human error, hemolysis, or concurrent analyses.
- Stated: findings apply to a population with serum NfL concentrations from 4.5 to 58.3 pg/mL and may not apply to higher concentrations. Evident from the text: the main cohort was small (n = 32) and diagnostically mixed, which may further limit broader generalizability.
Model-written summary. Check it against the paper before relying on it.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
3 of 6 cited sources checked · 3 supported · 0 flagged
- Interpretationown data
Inter-assay validity results are comparable to another large study investigating NfL and reporting assay precision.
“We provide data for inter-assay validity and found those results comparable to another large study investigating NfL and reporting assay precision15.”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [15] Khalil, M. et al. Serum neurofilament light levels in normal aging and their association with morphologic brain changes. Nat. Commun. 11, 812 (2020). source
- Findingown data
For samples stored for 7 days at room temperature, intraindividual variation (iCOV) is 9.0% for serum and 6.5% for plasma.
“For all three altered pre-freezing intervals, intraindividual variation (iCOV) was minimal (for samples stored for 7 days at room temperature; serum: 9.0%, plasma: 6.5%)”
Quote in the paperSubstantial claim - Findingown data
Inter-assay variability within four serum and four plasma samples measured on four consecutive runs was perfect.
“Inter-assay variability within four serum and four plasma samples measured on four consecutive runs was perfect.”
Quote in the paperNot a substantial claim - Findingown data
After 3 days at 4-8 °C, the mean absolute difference in NfL concentration compared to immediate freezing is 0.9 pg/mL for serum and 0.5 pg/mL for plasma.
“After storing of serum and blood tubes for 3 days at 4–8 °C, mean absolute differences compared to immediate freezing were 0.9 pg/mL for sNfL and 0.5 pg/mL for pNfL.”
Quote in the paperSubstantial claim - Findingown data
The mean concentration of NfL in plasma was 10% lower (95% CI 5-16) than in serum.
“First, the mean concentration of NfL in plasma was 10% lower (95% CI 5–16) than in serum which compares well to another investigation10.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [10] Hviid, C. V. B., Knudsen, C. S. & Parkner, T. Reference interval and preanalytical properties of serum neurofilament light chain in Scandinavian adults. Scand. J. Clin. Lab. Investig. https://doi.org/10.1080/00365513.2020.1730434 (2020). source
- Findingown data
NfL concentrations in serum and plasma remain stable at room temperature when processing of samples is delayed up to 7 days.
“we found concentrations of NfL in serum and plasma to remain stable at room temperature when processing of samples is delayed up to 7 days”
- Also stated in: § Discussion · plain
“we found that samples collected and stored at room temperature up to 7 days showed near-perfect stability of NfL concentrations in serum and a little less but still excellent stability in plasma”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Findingown data
After 7 days at room temperature, the mean absolute difference in NfL concentration compared to immediate freezing is 0.9 pg/mL for serum and 0.5 pg/mL for plasma.
“Ultimately, storing serum and blood vials for 7 days at room temperature resulted in a mean absolute difference in NfL concentrations compared to immediate freezing of 0.9 pg/mL for sNfL and 0.5 pg/mL for pNfL.”
Quote in the paperSubstantial claim - Findingown data
For samples stored for 7 days at room temperature, reproducibility is close to perfect with ICC = 0.99 for serum and ICC = 0.94 for plasma.
“reproducibility close to perfect (for samples stored for 7 days at room temperature; serum: ICC = 0.99, plasma: ICC = 0.94)”
- Also stated in: § Abstract · plain
“Consistency of these results was nearly perfect for serum and excellent for plasma (intraclass correlation coefficients 0.99 and 0.94, respectively).”
Quote in the paperSubstantial claim - Also stated in: § Abstract · plain
- Findingown data
Reliability for prolonged storage was slightly better in serum than plasma (ICC 0.99 vs. 0.94).
“Second, our results suggest that reliability for prolonged storage was slightly better in serum than plasma (ICC 0.99 vs. 0.94).”
Quote in the paperSubstantial claim - Findingown data
After 3 days at room temperature, the mean absolute difference in NfL concentration compared to immediate freezing is 1.4 pg/mL for serum and 0.8 pg/mL for plasma.
“Keeping samples stored for 3 days at room temperature yielded a mean absolute difference in concentration compared to immediate freezing of 1.4 pg/mL for sNfL and 0.8 pg/mL for pNfL.”
Quote in the paperSubstantial claim - Interpretationown data
Native serum samples may be slightly more suitable for shipment or any case of delayed processing for up to 7 days.
“Therefore, one may conclude that native serum samples may be slightly more suitable for shipment or any case of delayed processing for up to 7 days.”
Quote in the paperSubstantial claim - Interpretationcitation
The results obtained from our study align well within the manufacturer statements regarding the limits of detection.
“The results obtained from our study align well within the manufacturer statements regarding the limits of detection13.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [13] Quanterix. Data Sheets and Technical Notes for NF-light. https://www.quanterix.com/products-technology/assays/nf-lightr-sr-x-version (Accessed September 2020).
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- Interpretationown data
Native tubes can be stored or shipped for up to 7 days at room temperature and still yield reliable results for NfL concentrations.
“Native tubes can be stored or shipped for up to 7 days at room temperature and still yield reliable results for NfL concentrations.”
Quote in the paperSubstantial claim
- Classified as probably not a substantial claim. 12
- May combine two claims. 6
Classified as probably not a substantial claim. May combine two claims.
Study design and conductown dataAn a priori power calculation for detecting a systemic measurement error of 2 pg/mL (at 80% power and α-level 0.05) was performed, and 32 patients were included for the main study.
“Based on a previous study we performed an a priori power calculation for detecting a systemic measurement error of 2 pg/mL (at 80% power and an α-level of 0.05), we included groups consisting of (i) 32 patients for the main study of delayed processing of serum and plasma samples”
Quote in the paperSubstantial claim1 of 1 sources checked- cites [11] Altmann, P. et al. Serum neurofilament light chain withstands delayed freezing and repeated thawing. Sci. Rep. 10, 19982 (2020). source
May combine two claims.
InterpretationcitationOur results are in line with two other studies reporting pre-analytical stability of serum NfL in smaller cohorts, making this the largest study confirming this observation for both serum and plasma.
“Our results are in line with two other studies reporting pre-analytical stability of serum NfL in smaller cohorts making this the largest study confirming this observation for both serum and plasma10,11.”
Quote in the paperNot a substantial claim1 of 2 sources checked- cites [10] Hviid, C. V. B., Knudsen, C. S. & Parkner, T. Reference interval and preanalytical properties of serum neurofilament light chain in Scandinavian adults. Scand. J. Clin. Lab. Investig. https://doi.org/10.1080/00365513.2020.1730434 (2020). source
- cites [11] Altmann, P. et al. Serum neurofilament light chain withstands delayed freezing and repeated thawing. Sci. Rep. 10, 19982 (2020). source
May combine two claims.
Findingown dataSeparate analysis for the lowest and highest quartiles revealed still acceptable ICCs for sNfL and pNfL, though intraclass correlation was better for serum.
“Separate analysis for the lowest and highest quartiles revealed still acceptable ICCs for sNfL and pNfL though intraclass correlation was better for serum.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataFor sNfL, inter-assay iSD was 0.6 pg/mL (range 0.5-1.2), iCOV 4.8% (range 2.0-6.0), and ICC 1.0 (range 0.99-1.00).
“For sNfL, the iSD was 0.6 pg/mL (range 0.5–1.2), the iCOV 4.8% (range 2.0–6.0) and the ICC 1.0 (range 0.99–1.00).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown dataFor pNfL, inter-assay iSD was 1.6 pg/mL (range 0.8-2.7), iCOV 12.7% (range 6.0-18.0), and ICC 0.99 (range 0.90-1.00).
“For pNFL, the iSD was 1.6 pg/mL (range 0.8–2.7), the iCOV 12.7% (range 6.0–18.0) and the ICC 0.99 (range 0.90–1.00).”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenOnly samples yielding a coefficient of variance (CV) between paired replicates below 0.20 were included in this study.
“As per usual convention, only samples yielding a coefficient of variance (CV) between paired replicates below 0.20 were included in this study.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
InterpretationreasoningThe results may not be generalizable to a real-world busy lab setting due to greater risk of protocol deviation, human error, and other preanalytical errors.
“A real-world setting in a busy lab is expectedly more prone to accidental protocol deviation, human error, and other preanalytical errors such as hemolysis or the fact that various analyses are performed on a Quanterix reader for different analytes at the same time.”
Quote in the paperNot a substantial claim- Rests on an assumption: A real-world setting in a busy lab is more prone to accidental protocol deviation, human error, and other preanalytical errors such as hemolysis or simultaneous analyses on a Quanterix reader.
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductreasoningThis study complies with the World Medical Association Declaration of Helsinki regarding ethical conduct of research involving human subjects.
“This study complies with the World Medical Association Declaration of Helsinki regarding ethical conduct of research involving human subjects.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenThe ethics committee at the Medical University of Vienna approved this study (protocol number EK2100/2019, amended in July 2020).
“The ethics committee at the Medical University of Vienna (protocol number EK2100/2019, amended in July 2020) approved this study.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
InterpretationreasoningThe results may not be generalizable to other collection systems.
“Other collection systems may produce different results.”
Quote in the paperNot a substantial claim- Rests on an assumption: Other collection systems can produce different results.
May combine two claims.
Interpretationown dataThe results relate to a population showing sNfL concentrations ranging from 4.5 to 58.3 pg/mL and may not be applied to higher concentrations.
“Moreover, our results relate to a population showing sNfL concentrations ranging from 4.5 to 58.3 pg/mL and may not be applied to higher concentrations.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown dataInter-assay reproducibility was determined by repeatedly measuring NfL concentrations in 8 paired serum and plasma samples from 4 volunteers on four consecutive assay plates.
“In order to determine inter-assay reproducibility, we repeatedly measured NfL concentrations in 8 paired serum and plasma samples from 4 volunteers on four consecutive assay plates.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenInclusion of participants into the biobanking protocol followed written informed consent.
“Inclusion of participants into the biobanking protocol followed written informed consent.”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductreasoningOnly patients not taking any medication affecting thrombocyte function or blood coagulation were included.
“To avoid interference with our comparing measurements in serum and plasma, only patients not taking any medication affecting thrombocyte function of blood coagulation were included.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown dataNfL concentrations were analyzed by a blinded investigator.
“Serum and plasma aliquots were thawed for an hour at room temperature and analyzed by a blinded investigator.”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductnone givenWell and plate assignment was designed so that all aliquots from the same participant were analyzed in the same run to avoid within-subject run-to-run variability.
“Well and plate assignment was designed so that all aliquots from the same participant were analyzed in the same run to avoid within-subject run-to-run variability.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S2 (checkable), S4 (based).
Interpretationnone givenThe validity of the results depends on meticulous quality control in a specialized biomarker laboratory.
“Furthermore, the validity of our results depends on meticulous quality control in a specialized biomarker laboratory.”
Quote in the paperNot a substantial claim
The paper relies on this without support.
A real-world setting in a busy lab is more prone to accidental protocol deviation, human error, and other preanalytical errors such as hemolysis or simultaneous analyses on a Quanterix reader.
“A real-world setting in a busy lab is expectedly more prone to accidental protocol deviation, human error, and other preanalytical errors such as hemolysis or the fact that various analyses are performed on a Quanterix reader for different analytes at the same time.”
The paper relies on this without support.
Other collection systems can produce different results.
“Other collection systems may produce different results.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In those four studies, strengths of correlation were tested via Spearman's correlation coefficient.
“In all studies, however, strengths of correlation were tested via Spearman's correlation coefficient.”
From A Method to Combine Neurofilament Light Measurements From Blood Serum and Plasma in Clinical and Population-Based Studies (2022) · cites this paper as [18]
The Methods (Statistical analyses; page unavailable) say the paper calculated intraclass correlation coefficients (ICCs), not Spearman’s correlation coefficients. The Results (Inter-assay variability; page unavailable) likewise report ICCs for serum and plasma samples. (LLM, conf=0.87)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredSome studies report median differences between serum and plasma NfL and GFAP.
“Some studies report median differences,15–18 and only one has proposed a serum–plasma conversion for NfL in older adults, 19 with none for GFAP.”
From Establishing demographic-based normative values for neurofilament light chain and glial fibrillary acidic protein in serum and plasma samples (2026) · cites this paper as [17]
The Discussion reports a serum–plasma difference for NfL: mean plasma concentration was 10% lower than serum concentration. It does not report a median difference or address GFAP. The later claim about conversion is attached to a different citation. (LLM, conf=0.86)
Experimental — accuracy not yet measuredFour studies consistently reported higher NfL levels in serum than in plasma.
“They consistently reported higher NfL levels in serum than in plasma.”
From A Method to Combine Neurofilament Light Measurements From Blood Serum and Plasma in Clinical and Population-Based Studies (2022) · cites this paper as [18]
We confirm the numerical differences between serum and EDTA-plasma NfL levels that have been found in other studies.
“We confirm the numerical differences between serum and EDTA-plasma NfL levels that have been found in other studies (15–18).”
From A Method to Combine Neurofilament Light Measurements From Blood Serum and Plasma in Clinical and Population-Based Studies (2022) · cites this paper as [18]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
This check didn’t finish.
Cites
Rissin, D. M., Kan, C. W., Campbell, T. G., Howes, S. C., Fournier, D. R., Song, L., Piech, T., Patel, P. P., Chang, L., Rivnak, A. J., et al. (2010) · Nature Biotechnology · doi:10.1038/nbt.1641 · Cited by 2,119 · see citing works
Neurofilaments as biomarkers in neurological disorders
Khalil, M., Teunissen, C. E., Otto, M., Piehl, F., Sormani, M. P., Gattringer, T., Barro, C., Kappos, L., Comabella, M., Fazekas, F., et al. (2018) · Nature Reviews Neurology · doi:10.1038/s41582-018-0058-z · Cited by 1,988 · see citing works
Self-citationNeurofilament light chain as a biomarker in neurological disorders
Gaetani, L., Blennow, K., Calabresi, P., Di Filippo, M., Parnetti, L., & Zetterberg, H. (2019) · Journal of Neurology, Neurosurgery & Psychiatry · doi:10.1136/jnnp-2018-320106 · Cited by 1,334 · see citing works
Self-citationSerum neurofilament light: a biomarker of neuronal damage in multiple sclerosis
Disanto, G., Barro, C., Benkert, P., Naegelin, Y., Schädelin, S., Giardiello, A., Zecca, C., Blennow, K., Zetterberg, H., Leppert, D., et al. (2017) · Annals of Neurology · doi:10.1002/ana.24954 · Cited by 1,164 · see citing works
Self-citationClinical biostatistics: LIV. The biostatistics of concordance
Kramer, M. S., & Feinstein, A. R. (1981) · Clinical Pharmacology & Therapeutics · doi:10.1038/clpt.1981.18 · Cited by 1,087 · see citing works
Kuhle, J., Barro, C., Andreasson, U., Derfuss, T., Lindberg, R., Sandelius, Å., Liman, V., Norgren, N., Blennow, K., & Zetterberg, H. (2016) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2015-1195 · Cited by 740 · see citing works
Self-citationKhalil, M., Pirpamer, L., Hofer, E., Voortman, M. M., Barro, C., Leppert, D., Benkert, P., Ropele, S., Enzinger, C., Fazekas, F., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-14612-6 · Cited by 572 · see citing works
Kuhle, J., Barro, C., Disanto, G., Mathias, A., Soneson, C., Bonnier, G., Yaldizli, Ö., Regeniter, A., Derfuss, T., Canales, M., et al. (2016) · Multiple Sclerosis Journal · doi:10.1177/1352458515623365 · Cited by 257 · see citing works
Neurofilament Light: A Dynamic Cross-Disease Fluid Biomarker for Neurodegeneration
Zetterberg, H. (2016) · Neuron · doi:10.1016/j.neuron.2016.06.030 · Cited by 197 · see citing works
Self-citationHviid, C. V. B., Knudsen, C. S., & Parkner, T. (2020) · Scandinavian Journal of Clinical and Laboratory Investigation · doi:10.1080/00365513.2020.1730434 · Cited by 143 · see citing works
Haslacher, H., Gerner, M. C., Hofer, P., Jurkowitsch, A., Hainfellner, J. A., Kain, R., Wagner, O. F., & Perkmann, T. (2018) · Biopreservation and Biobanking · doi:10.1089/bio.2018.0032 · Cited by 96 · see citing works
Self-citationAltmann, P., Simoni, D. D., Kaider, A., Ludwig, B., Rath, J., Leutmezer, F., Zimprich, F., Höftberger, R., Lunn, M. P., Heslegrave, A., et al. (2020) · Journal of Neuroinflammation · doi:10.1186/s12974-020-01737-0 · Cited by 81 · see citing works
Self-citationSerum neurofilament light chain withstands delayed freezing and repeated thawing
Altmann, P., Leutmezer, F., Zach, H., Wurm, R., Stattmann, M., Ponleitner, M., Petzold, A., Zetterberg, H., Berger, T., Rommer, P., et al. (2020) · Scientific Reports · doi:10.1038/s41598-020-77098-8 · Cited by 67 · see citing works
Self-citationNeurofilaments: The C-Reactive Protein of Neurology
Lambertsen, K. L., Soares, C. B., Gaist, D., & Nielsen, H. H. (2020) · Brain Sciences · doi:10.3390/brainsci10010056 · Cited by 66 · see citing works
Serum neurofilament levels correlate with retinal nerve fiber layer thinning in multiple sclerosis
Bsteh, G., Berek, K., Hegen, H., Teuchner, B., Buchmann, A., Voortman, M. M., Auer, M., Wurth, S., Zinganell, A., Pauli, F. D., et al. (2019) · Multiple Sclerosis Journal · doi:10.1177/1352458519882279 · Cited by 37 · see citing works
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Neurofilaments as biomarkers in neurological disorders — towards clinical application
Khalil, M., Teunissen, C. E., Lehmann, S., Otto, M., Piehl, F., Ziemssen, T., Bittner, S., Sormani, M. P., Gattringer, T., Abu‐Rumeileh, S., et al. (2024) · Nature Reviews Neurology · doi:10.1038/s41582-024-00955-x · Cited by 484 · see citing works
“This proteolytic breakdown of neurofilaments yields highly soluble and stable cleavage products 2,17,47 .”
Self-citationArslan, B., & Zetterberg, H. (2023) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2023-0036 · Cited by 69 · see citing works
Self-citationPlasma biomarkers for neurodegenerative disorders: ready for prime time?
Balogun, W. G., Zetterberg, H., Blennow, K., & Karikari, T. K. (2023) · Current Opinion in Psychiatry · doi:10.1097/yco.0000000000000851 · Cited by 59 · see citing works
Self-citationvan Lierop, Z. Y., Verberk, I. M. W., van Uffelen, K. W. J., Simmelink, M. J. A., Veld, L. I. ‘., Killestein, J., & Teunissen, C. E. (2022) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2022-0007 · Cited by 53 · see citing works
The 2022 Lady Estelle Wolfson lectureship on neurofilaments
Petzold, A. (2022) · Journal of Neurochemistry · doi:10.1111/jnc.15682 · Cited by 49 · see citing works
“It was the serendipitous finding of very long- term stability of NfL from an ancient human brain (Petzold et al., 2020) which stimulated re- visitation of the question in a joint experiment (Altmann et al., 2021).”
BackgroundMethodInfluentialSelf-citationBornhorst, J. A., Figdore, D. J., Campbell, M. R., Pazdernik, V., Mielke, M. M., Petersen, R. C., & Algeciras‐Schimnich, A. (2022) · Clinica Chimica Acta · doi:10.1016/j.cca.2022.08.017 · Cited by 50 · see citing works
Blood Biomarkers for Alzheimer’s Disease in Down Syndrome
Montoliu‐Gaya, L., Strydom, A., Blennow, K., Zetterberg, H., & Ashton, N. J. (2021) · Journal of Clinical Medicine · doi:10.3390/jcm10163639 · Cited by 45 · see citing works
Self-citationAssociations of sNfL with clinico‐radiological measures in a large MS population
Sotirchos, E. S., Fitzgerald, K. C., Singh, C. M., Smith, M. D., Reyes-Mantilla, M. I., Hersh, C. M., Hyland, M. H., Canissario, R., Simmons, S. B., Arrambide, G., et al. (2022) · Annals of Clinical and Translational Neurology · doi:10.1002/acn3.51704 · Cited by 45 · see citing works
“This observation is unlikely to be related to a technical issue due to different collection procedures and/or processing, given that sNfL levels have been previously reported to remain stable in unprocessed blood samples stored up to 7 days at room temperature, to withstand repeat freeze–thaw cycles, and to be similar…”
ResultYoussef, P., Hughes, L., Kim, W. S., Halliday, G. M., Lewis, S. J. G., Cooper, A., & Dzamko, N. (2023) · Scientific Reports · doi:10.1038/s41598-023-32480-0 · Cited by 46 · see citing works
Bircak-Kuchtova, B., Chung, H., Wickel, J., Ehler, J., & Geis, C. (2023) · Critical Care · doi:10.1186/s13054-023-04497-4 · Cited by 46 · see citing works
Wieske, L., Smyth, D., Lunn, M. P., Eftimov, F., & Teunissen, C. E. (2021) · Neurotherapeutics · doi:10.1007/s13311-021-01136-0 · Cited by 35 · see citing works
“Serum and plasma NfL are both stable at room temperature for up to 7 days, and NfL is also stable to up to 4 freeze–thaw cycles and centrifugations [41, 43].”
BackgroundShahim, P., Norato, G., Sinaii, N., Zetterberg, H., Blennow, K., Chan, L. E., & Grunseich, C. (2024) · Genes · doi:10.3390/genes15040496 · Cited by 37 · see citing works
“In comparison, a previous study [51] measuring NfL in samples of patients with various neurologic conditions found that the NfL level is between 10 and 12% higher in serum than in plasma.”
ResultSelf-citationJing, X., Wang, L., Song, M. L., Geng, H., Li, W., Huo, Y., Huang, A., Wang, X., & An, C. (2024) · Scientific Reports · doi:10.1038/s41598-024-59530-5 · Cited by 35 · see citing works
Pre-Analytical Variables Influencing Stability of Blood-Based Biomarkers of Neuropathology
Panikkar, D., Vivek, S., Crimmins, E. M., Faul, J. D., Langa, K. M., & Thyagarajan, B. (2023) · Journal of Alzheimer s Disease · doi:10.3233/jad-230384 · Cited by 34 · see citing works
Neurofilament light chain as a biomarker of chemotherapy-induced peripheral neuropathy
Andersen, N. E., Boehmerle, W., Huehnchen, P., et al. (2024) · Trends in Pharmacological Sciences · doi:10.1016/j.tips.2024.08.001 · Cited by 30 · see citing works
Kapoor, M., Carr, A. S., Foiani, M. S., et al. (2022) · European Journal of Neurology · doi:10.1111/ene.15496 · Cited by 29 · see citing works
Self-citationRübsamen, N., Willemse, E. A. J., Leppert, D., Wiendl, H., Nauck, M., Karch, A., Kuhle, J., & Berger, K. (2022) · Frontiers in Neurology · doi:10.3389/fneur.2022.894119 · Cited by 27 · see citing works
“(18) report that reliability for prolonged storage was slightly better in serum than EDTA-plasma and conclude that native serum samples may be slightly more suitable for shipment or any case of delayed processing.”
BackgroundThe clinical and molecular spectrum of ZFYVE26-associated hereditary spastic paraplegia: SPG15
Saffari, A., Kellner, M., Jordan, C., et al. (2022) · Brain · doi:10.1093/brain/awac391 · Cited by 27 · see citing works
Wilson, D. H., Chan, D., Chang, L., Mathis, R., Verberk, I. M. W., Montalbán, X., Comabella, M., Fissolo, N., Bielekova, B., Masvekar, R., et al. (2023) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2023-0518 · Cited by 26 · see citing works
Self-citationVietzen, H., Kühner, L. M., Berger, S., Ponleitner, M., Graninger, M., Pistorius, C., Jungbauer, C., Reindl, M., Saucke, H., Kauth, F., et al. (2025) · Nature Communications · doi:10.1038/s41467-025-61751-9 · Cited by 24 · see citing works
Self-citationGonzalez‐Robles, C., Weil, R. S., van Wamelen, D. J., Bartlett, M., Burnell, M., Clarke, C. S., Hu, M. T., Huxford, B., Jha, A., Lambert, C., et al. (2023) · Journal of Parkinson s Disease · doi:10.3233/jpd-230051 · Cited by 23 · see citing works
Self-citationThe Biochemical Assessment of Mitochondrial Respiratory Chain Disorders
Turton, N., Cufflin, N., Dewsbury, M., et al. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23137487 · Cited by 21 · see citing works
Abdelhak, A., Bachhuber, F., Ning, K., Benkert, P., Boscardin, W. J., Maceski, A. M., Schaedelin, S. A., Achtnichts, L., Finkener, S., Lalive, P. H., et al. (2025) · Journal of Neurology Neurosurgery & Psychiatry · doi:10.1136/jnnp-2025-335831 · Cited by 20 · see citing works
“Our large multinational collaboration harnesses the power of IPD to substantiate the growing evidence of the prognostic value of GFAP and NfL in PMS. 24–27 All those strengths render both markers promising candidates in MS research and for application in MS care.”
InfluentialBlood Biomarkers for Return to Play after Concussion in Professional Rugby Players
Oris, C., Durif, J., Rouzaire, M., Pereira, B., Bouvier, D., Kahouadji, S., Abbot, M., Braïlova, M., Lehmann, S., Hirtz, C., et al. (2022) · Journal of Neurotrauma · doi:10.1089/neu.2022.0148 · Cited by 19 · see citing works
“…presents a pre-analytical advantage due to its relative stability, at least 8 h at room temperature or 48 h at 2-8 (cid:2) C. 23 NFL shows a similar robustness in terms of pre-analytical characteristics, 24-26 but differs analytically with a higher CV% (intra-assay CV: 6.6%; inter-assay CV: 17%).”
InfluentialSchmitz, M., Canaslan, S., Espinosa, J. C., Fernández‐Borges, N., Villar‐Piqué, A., Llorens, F., Varges, D., Maass, F., Torres, J. M., Hermann, P., et al. (2022) · Molecular Neurobiology · doi:10.1007/s12035-022-02891-7 · Cited by 18 · see citing works
Neurofilament Light Chains in Systemic Amyloidosis: A Systematic Review
Berends, M., Nienhuis, H. L. A., Adams, D., et al. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25073770 · Cited by 18 · see citing works
Markers of axonal injury in blood and tissue triggered by acute and chronic demyelination
Abdelhak, A., Cordano, C., Duncan, G. J., Emberley, K., Nocera, S., Xin, W., Ananth, K., Jabassini, N., Ning, K., Reinsberg, H., et al. (2025) · Brain · doi:10.1093/brain/awaf144 · Cited by 17 · see citing works
Self-citationSotirchos, E. S., Hu, C., Smith, M. D., Lord, H., DuVal, A. L., Arrambide, G., Montalbán, X., Akgün, K., Ziemssen, T., Naismith, R. T., et al. (2023) · Neurology · doi:10.1212/wnl.0000000000207957 · Cited by 16 · see citing works
Fridman, V., Sillau, S., Ritchie, A. M., Bockhorst, J., Coughlan, C. M., Araya, P., Espinosa, J. M., Smith, K. P., Lange, E. M., Lange, L. A., et al. (2023) · Journal of the Peripheral Nervous System · doi:10.1111/jns.12575 · Cited by 15 · see citing works
“Among candidate plasma biomarkers for peripheral neuropathy, NFL has several notable advantages, including high concentrations in axons, availability in biofluids,(15) and stability in stored samples.(16) We performed a case– control study evaluating plasma NFL concentrations over a 12-year period in participants of t…”
Comparing Neurofilament Light Chain Levels in Serum and Plasma
Kwon, H. S., Lee, H., Kim, Y. S., Choi, H., Lee, K.-Y., Lee, Y. J., Lee, E.-H., Hwang, M., Park, H., & Koh, S.-H. (2023) · Dementia and Neurocognitive Disorders · doi:10.12779/dnd.2023.22.3.109 · Cited by 15 · see citing works
Chen, Y., Zeng, X., Diaz, J. L., Sehrawat, A., Lafferty, T. K., Boslett, J. J., Klunk, W. E., Pascoal, T. A., Villemagne, V. L., Cohen, A. D., et al. (2024) · Journal of Neurochemistry · doi:10.1111/jnc.16130 · Cited by 15 · see citing works
“All three biomarkers remained stable at room temperature, which is consistent with previous results (Altmann et al., 2021 ; Ashton, Suárez-Calvet, et al., 2021 ; Bowen et al., 2010; Verberk et al., 2022 ).”
Nicolella, V., Fiorenza, M., Monteiro, I., et al. (2024) · Journal of the Neurological Sciences · doi:10.1016/j.jns.2024.123115 · Cited by 14 · see citing works
“Very few studies have compared serum and plasma measurements of NfL and have reported conflicting results [30 – 32].”
Amrein, M., Meier, S., Schäfer, I., Schaedelin, S. A., Willemse, E. A. J., Benkert, P., Walter, J. E., Puelacher, C., Zimmermann, T., Median, D., et al. (2023) · Biomarkers · doi:10.1080/1354750x.2023.2172211 · Cited by 13 · see citing works
Alecu, J. E., Saffari, A., Ziegler, M., et al. (2023) · Movement Disorders · doi:10.1002/mds.29524 · Cited by 12 · see citing works
Mondésert, E., Schraen‐Maschke, S., Quadrio, I., Bousiges, O., Bouvier, D., Delaby, C., Bedel, A., Lehmann, S., & Fourier, A. (2024) · Clinica Chimica Acta · doi:10.1016/j.cca.2024.120007 · Cited by 13 · see citing works
Rosadas, C., & Taylor, G. P. (2023) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2022-0904 · Cited by 10 · see citing works
Schaller, M., Maiworm, M., Schäfer, J. H., Friedauer, L., Hattingen, E., Wenger, K. J., Weber, F., Jakob, J., Steffen, F., Bittner, S., et al. (2024) · Journal of Neurology · doi:10.1007/s00415-024-12299-z · Cited by 9 · see citing works
Neurofilament Light Chain as Biomarker in Encephalitis
Wellmann, S. M., Geis, T., Kühle, J., et al. (2024) · Journal of Clinical Medicine · doi:10.3390/jcm13185416 · Cited by 9 · see citing works
“…and in both serum and plasma, NfL has been proven to be very stable with unchanged concentrations over a period of seven days at room temperature [4]. three to four weeks, and in both serum and plasma, NfL has been proven to be very stable with unchanged concentrations over a period of seven…”
BackgroundMäättä, L. L., Andersen, S. T., Parkner, T., Hviid, C. V. B., Witte, D. R., John, J., Pascal, M. M. V., Ferris, E., Baskozos, G., Ramirez, J. D., et al. (2025) · Diabetes Research and Clinical Practice · doi:10.1016/j.diabres.2025.112098 · Cited by 8 · see citing works
Neurofilament heavy phosphorylated epitopes as biomarkers in ageing and neurodegenerative disease
Paoli, L. F. D., Kirkcaldie, M., King, A. E., & Collins, J. M. (2024) · Journal of Neurochemistry · doi:10.1111/jnc.16261 · Cited by 7 · see citing works
“Second, the form of NFL detected in biomarker assays is shown to be very stable in blood samples (Altmann et al., 2021), and while its half-life in the blood is unknown, one study has estimated it to be 43 days in serum (Mortensen et al., 2023 ).”
Blood Biomarkers Reflecting Brain Pathology—From Common Grounds to Rare Frontiers
Weinhofer, I., Rommer, P. S., & Berger, J. (2025) · Journal of Inherited Metabolic Disease · doi:10.1002/jimd.70032 · Cited by 8 · see citing works
Self-citationUse of neurofilament light chain to identify structural brain diseases in dogs
Sung, J., Chae, Y., Yun, T., Koo, Y., Lee, D., Kim, H., Yang, M., & Kang, B. (2024) · Journal of Veterinary Internal Medicine · doi:10.1111/jvim.17110 · Cited by 8 · see citing works
Krajnc, N., Altmann, P., Riedl, K., Mitsch, C., Berger, T., Leutmezer, F., Rommer, P. S., Pemp, B., & Bsteh, G. (2022) · Frontiers in Neurology · doi:10.3389/fneur.2022.814734 · Cited by 6 · see citing works
“Neurofilament Light Chains Serum and CSF samples were extracted from our neurological biobank where aliquots are stored at −70C in accordance with international consensus guidelines (24, 25).”
MethodSelf-citationColeman, A., Touzé, A., Farag, M., Pengo, M., Murphy, M., Hassan, Y., Thackeray, O., Fayer, K., Field, S., Nakajima, M., et al. (2025) · Journal of Neurology · doi:10.1007/s00415-025-13232-8 · Cited by 9 · see citing works
“Several studies have demonstrated that NfL is highly stable in venous blood plasma and can withstand freeze-thawing and delayed sample processing, even up to seven days [21, 22].”
Self-citationMeter, T. E. V., Wilson, E. N., Elahi, F. M., et al. (2026) · Alzheimer s & Dementia · doi:10.1002/alz.71252 · Cited by 5 · see citing works
Streit, S., Meinhardt, J., Gimber, N., Kestenbach, J., Siewert, C., Schmoranzer, J., Meisel, C., Ruprecht, K., Heppner, F. L., Körtvélyessy, P., et al. (2025) · Brain Pathology · doi:10.1111/bpa.70038 · Cited by 5 · see citing works
“However, NfL-levels in serum and CSF are very stable both when cryoarchived [36 – 39] and at room temperature [38 – 40], and the median time in our cohort between collection of serum/CSF and biopsy was only 4 days.”
Dilixiati, D., Kadier, K., Qiao, B., Zhang, W., Nuerdebieke, D., Zebibula, A., Yang, Y., & Rexiati, M. (2025) · BMJ Open · doi:10.1136/bmjopen-2024-084766 · Cited by 4 · see citing works
“However, further high-quality prospective cohort studies are needed to verify the causal relationship between elevated serum NfL levels and reduced grip strength.”
Type of serum collection tube does not impact neurofilament light chain levels
Sotirchos, E. S., Fitzgerald, K. C., Smith, M. D., Vasileiou, E. S., Resto, Y., Lord, H., Mowry, E. M., & Calabresi, P. A. (2022) · Multiple Sclerosis and Related Disorders · doi:10.1016/j.msard.2022.103676 · Cited by 4 · see citing works
Neurofilament Light Chain Is Associated With Acute Mountain Sickness
Berek, K., Lindner, A., Pauli, F. D., Bsteh, G., Treml, B., Ponleitner, M., Engler, C., Kleinsasser, A. T., Berger, T., Wille, M., et al. (2024) · Brain and Behavior · doi:10.1002/brb3.70165 · Cited by 3 · see citing works
“Some of the low effect size may be explained by our study using EDTA plasma samples, since it is known that absolute levels of NfL are lower if measured in plasma than in serum (Altmann et al. 2021).”
Self-citationHanin, A., Bahroun, S., Chiodega, V., Bonnefont‐Rousselot, D., Amador, M. D., Salachas, F., Mariotto, S., Dingeo, G., & Lamari, F. (2026) · Revue Neurologique · doi:10.1016/j.neurol.2026.03.013 · Cited by 2 · see citing works
Rojas-Núñez, I., Gomez, A. M. M., Palmer, S. E., et al. (2022) · Journal of Equine Veterinary Science · doi:10.1016/j.jevs.2022.104057 · Cited by 2 · see citing works
Proteolysis‐Based Biomarker Repertoire of the Neurofilament Proteome
Petzold, A. (2025) · Journal of Neurochemistry · doi:10.1111/jnc.70023 · Cited by 2 · see citing works
“Such selection can be refined by focusing on the thermodynamically most stable and soluble peptides, a prerequisite for any successful laboratory biomarker (Petzold 2022; Altmann et al. 2021; Khalil et al. 2024).”
Self-citationBorko, T. L., Ritchie, A., Selva, S., Sillau, S., Engebretson, E., Wolf, A. B., Alvarez, E., Gross, R., Gutierrez, E., Corboy, J., et al. (2026) · Multiple Sclerosis Journal – Experimental, Translational and Clinical · doi:10.1177/20552173261466440 · Cited by 1 · see citing works
Perino, J. H., Patterson, M., Momen, M. M., et al. (2024) · American Journal of Veterinary Research · doi:10.2460/ajvr.23.12.0292 · Cited by 1 · see citing works
“9 All 4 subunits act together to aid in the growth of axonal diameter and act as axonal scaffolding.”
Self-citationBrown, Q. L., Nicholson, E. M., Wang, C., Greenlee, J. J., Seger, H., Veneziano, S. E., & Cassmann, E. D. (2024) · PLoS ONE · doi:10.1371/journal.pone.0299038 · Cited by 1 · see citing works
Choucair, I., Lee, T., Eldik, L. J. V., Dage, J. L., Gold, B. T., Abner, E. L., Jicha, G. A., & Nelson, P. T. (2025) · Journal of Neuropathology & Experimental Neurology · doi:10.1093/jnen/nlaf115 · Cited by 2 · see citing works
“…our findings with NfL contradicted the studies that used − 80 � C storage to lock-in experimental condition and batch all samples together 11,18,21 and with the Roche study showing NfL stability at room temperature for 24 h. 16 In contrast to NfL, detected GFAP levels exhibited relatively…”
SLC7A11 and NLRP1 in non-small cell lung cancer
Khan, A., Alzahrani, A. R., Jawaid, T., et al. (2025) · Clinica Chimica Acta · doi:10.1016/j.cca.2025.120684 · Cited by 1 · see citing works
Wellmann, S. M., & Lehnerer, V. (2024) · Trillium Diagnostik · doi:10.47184/td.2024.02.07
Hochwaldstrasse 2, D.-9. B. B., & Goischke, H.-K. (2025) · Neurology and Neuroscience · doi:10.33425/2692-7918.1084
Fluid Markers of Axonal and Neuronal Injury
Arslan, B., & Zetterberg, H. (2025) · Springer eBooks · doi:10.1007/978-3-031-80849-4_5
Self-citationDargvainiene, J., Helmers, A., Naumann, J., Gless, C. A., Möller, B., Welzel, J., Leypoldt, F., Hensler, J., Tesseur, I. C., Wandinger, K., et al. (2025) · Movement Disorders · doi:10.1002/mds.70071
Bergmann, C., Häsler, L. M., Lambert, M., Kaeser, S. A., Schultz, S. A., Riond, B., Weiss, M., Balz, M., Knauf‐Witzens, T., & Jucker, M. (2026) · PLoS Biology · doi:10.1371/journal.pbio.3003606
“Previous research has found that human serum NfL remains stable at least up to 7 days at room temperature and withstands at least 3 freeze-thaw cycles [31,32].”
Weaver, R. D., & Lei, M. (2026) · Alzheimer s & Dementia Behavior & Socioeconomics of Aging · doi:10.1002/bsa3.70069
Rajesh, N., Jasmine, R., Joji, R., Sivakumar, S., & Francis, D. (2026) · International review of neurobiology · doi:10.1016/bs.irn.2026.05.009
Škorvanová, M., Verespejova, A. I., Hanes, J., et al. (2026) · PLoS ONE · doi:10.1371/journal.pone.0357207
- Authors
- Altmann, P., Ponleitner, M., Rommer, P. S., Haslacher, H., Mucher, P., Leutmezer, F., Petzold, A., Wotawa, C., Lanzenberger, R., Berger, T., Zetterberg, H., & Bsteh, G.
- Affiliations
- Medical University of Vienna (AT)
- UCL Queen Square Institute of Neurology (GB)
- Sahlgrenska University Hospital (SE)
- Corresponding author
- Bsteh, Gabriel
- ORCID
- Venue
- Scientific Reports
- Published
- 2021-05-26
- Volume
- 11
- Issue
- 1
- Pages
- 11034-11034
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC8154890
- OpenAlex
- W3165219122
- Semantic Scholar
- 5fe0478ee10412a89bbb709e65053ba1f6a129ef
- Cited by
- 68
- References
- 15
- Citations by year
- 2026: 7 · 2025: 14 · 2024: 16 · 2023: 15 · 2022: 13 · 2021: 3
- FWCI
- 6.11
- Type
- journal article
- Language
- en
- Update notices
- correction (2022-04-20)
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Amsterdam UMC (VU Amsterdam) - Institutional Repository (publishedVersion, other-oa)
- Pure Amsterdam UMC (acceptedVersion, cc-by)
- PubMed Central (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- UCL Discovery (University College London) (submittedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion, other-oa)
- Funders
- Universität Wien
- Medizinische Universität Wien
- Topics
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Hereditary Neurological Disorders
- Keywords
- neurofilament light chain, pre-analytical stability, serum, plasma, room temperature storage, sample storage, Simoa, neurological disorders, sample shipment
- MeSH terms
- Adult, Aged, Female, Humans, Immunoassay, Intermediate Filaments, Male, Middle Aged, Nervous System Diseases, Plasma, Specimen Handling, Time Factors, Biomarkers, Neurofilament Proteins, Protein Stability
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-17.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-17.
Cited sources are checked by gpt-6-sol.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
