Nf light assay

Manufactured by Quanterix

Sourced in United States

The NF-Light assay is a quantitative immunoassay developed by Quanterix to measure levels of Neurofilament Light (NF-L), a biomarker associated with neurological conditions. The assay leverages Quanterix's proprietary Single Molecule Array (Simoa) technology to provide highly sensitive and accurate detection of NF-L in biological samples.

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Close spelling variants of this product

NF-light” assay kits Simoa NF-light assay Simoa™ NF-light® Advantage assay NF-Light Simoa Assay Advantage kit NF-Light

8 protocols using nf light assay

Serum Neurofilament Light Quantification

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A subgroup of patients (n = 192) underwent blood collection for serum NfL measurement; 63 healthy controls (HC) (age 67.0, IQR 61.0–74.0 years) were recruited among spouses or caregivers as reference.
Serum was collected by venipuncture, processed and stored in aliquots at – 80 °C according to standardized procedures. Serum NfL was measured by single-molecule array (Simoa) technology on an HD-X Analyzer using the commercial NF-light® assay according to the manufacturer’s instructions (Quanterix, Billerica, MA). Detailed analytical procedures and assay validation have been previously described [15 (link)]. The lower limits of quantitation for serum NfL were 0.174 pg/mL. The measurements were performed out in one round of experiments using the same batch of reagents, and the operators were blinded to all clinical information. Quality control samples had intra-assay and inter-assay coefficients of variation of less than 8 and 20%, respectively.

Benussi A., Ashton N.J., Karikari T.K., Alberici A., Saraceno C., Ghidoni R., Benussi L., Zetterberg H., Blennow K, & Borroni B. (2021). Prodromal frontotemporal dementia: clinical features and predictors of progression. Alzheimer's Research & Therapy, 13, 188.

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Quantification of Neurodegeneration Biomarkers

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A subgroup of patients (n = 199) underwent blood collection for serum neurofilament light (NfL) and serum p‐tau₁₈₁ dosages by single molecule array (Simoa) technology, as previously described.²⁰, ²¹ Briefly, NfL were measured on an HD‐X Analyzer using the commercial NF‐Light^® assay according to the manufacturer's instructions (Quanterix=) with lower limit of quantitation of 0.174 pg/mL. Serum p‐tau₁₈₁ was measured using an in‐house Simoa assay developed at the University of Gothenburg with the lower limit of quantitation of 1 pg/mL. The measurements were performed in one round of experiments using the same batch of reagents, and the operators were blinded to all clinical information. Quality control samples had intra‐assay and inter‐assay coefficients of variation of less than 8% and 20%, respectively.

Benussi A., Libri I., Premi E., Alberici A., Cantoni V., Gadola Y., Rivolta J., Pengo M., Gazzina S., Calhoun V.D., Gasparotti R., Zetterberg H., Ashton N.J., Blennow K., Padovani A, & Borroni B. (2022). Differences and similarities between familial and sporadic frontotemporal dementia: An Italian single‐center cohort study. Alzheimer's & Dementia : Translational Research & Clinical Interventions, 8(1), e12326.

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Quantifying Serum Neurofilament Light

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sNfL was measured (at baseline and every 6 or 12 months) with the NF‐light® assay (Quanterix, Billerica, MA, USA).
Intra‐ and interassay variability were evaluated with 3 native serum samples in each of the runs on independent days. The mean coefficients of variation (CVs) of duplicate determinations for concentration were 5.5% (6.2pg/ml, sample 1), 3.3% (18.9pg/ml, sample 2), and 3.1% (37.3pg/ml, sample 3). Interassay CVs were 7.0% (sample 1), 5.6% (sample 2), and 5.6% (sample 3). For the generation of age‐dependent sNfL Z‐scores, we used sNfL measurements in 8,865 samples of 4,209 healthy controls (median age = 44.8 years).²¹

Oechtering J., Schaedelin S., Benkert P., Müller S., Achtnichts L., Vehoff J., Disanto G., Findling O., Fischer‐Barnicol B., Orleth A., Chan A., Pot C., Barakovic M., Rahmanzadeh R., Galbusera R., Heijnen I., Lalive P.H., Wuerfel J., Subramaniam S., Aeschbacher S., Conen D., Naegelin Y., Maceski A., Meier S., Berger K., Wiendl H., Lincke T., Lieb J., Yaldizli Ö., Sinnecker T., Derfuss T., Regeniter A., Zecca C., Gobbi C., Kappos L., Granziera C., Leppert D, & Kuhle J. (2021). Intrathecal Immunoglobulin M Synthesis is an Independent Biomarker for Higher Disease Activity and Severity in Multiple Sclerosis. Annals of Neurology, 90(3), 477-489.

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Quantification of Neurofilament Light in Serum

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In a subgroup of patients (n = 188), the serum was collected to assess the concentrations of NfL. The serum was obtained by venipuncture, processed and stored in aliquots at − 80 °C according to standardized procedures. The serum NfL concentration was measured using a commercial NF-Light assay (Quanterix, Billerica, Massachusetts, USA) according to the manufacturer’s instructions. The lower limit of quantitation for serum NfL was 0.174 pg/ml. Measurements were carried out using a HD-X analyser (Quanterix, Billerica, Massachusetts, USA), and the operators were blinded to all clinical information. Quality control samples had mean intra- and inter-assay coefficients of variation of < 8% and < 20%, respectively.

Pengo M., Alberici A., Libri I., Benussi A., Gadola Y., Ashton N.J., Zetterberg H., Blennow K, & Borroni B. (2022). Sex influences clinical phenotype in frontotemporal dementia. Neurological Sciences, 43(9), 5281-5287.

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Biomarkers for Neurodegenerative Diseases

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All serum and CSF samples were centralized at the Clinical Proteomics Platform of the Laboratory of Biochemistry—Clinical Proteomic in Montpellier. sNfL and cNfL concentrations were determined using commercial NF-Light assay (Quanterix, Billerica, MA) based on ultrasensitive Simoa technology, as previously described.^{28 (link)} CSF CHI3L1 levels were measured by ELISA using the MicroVue YKL-40 EIA kit (Quidel Corporation, San Diego, CA), as already described.^{29 (link)} All experiments were performed with a single batch of reagents by researchers blinded from any clinical information. There were no missing values for NfL or CHI3L1 measures.

Rival M., Thouvenot E., Du Trieu de Terdonck L., Laurent-Chabalier S., Demattei C., Uygunoglu U., Castelnovo G., Cohen M., Okuda D.T., Kantarci O.H., Pelletier D., Azevedo C., Marin P., Lehmann S., Siva A., Mura T, & Lebrun-Frenay C. (2022). Neurofilament Light Chain Levels Are Predictive of Clinical Conversion in Radiologically Isolated Syndrome. Neurology® Neuroimmunology & Neuroinflammation, 10(1), e200044.

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CSF NfL Quantification Protocol

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Chain Levels CSF NfL was measured in duplicate with the NF-light ® assay (Quanterix, Billerica, MA). The mean intra-assay coefficient of variation (CV) of duplicate determinations for CSF NfL was 2.5%. Few samples with intra-assay CVs >20% were repeatedly measured. The mean interassay CVs of internal quality controls for NfL were 6.8% (8.7 pg/mL), 4.4% (17.9 pg/mL), and 6.1% (106.5 pg/mL).

Oechtering J., Stein K., Schaedelin S.A., Maceski A.M., Orleth A., Meier S., Willemse E., Qureshi F., Heijnen I., Regeniter A., Derfuss T., Benkert P., D'Souza M., Limberg M., Fischer-Barnicol B., Achtnichts L., Mueller S., Salmen A., Lalive P.H., Bridel C., Pot C., Du Pasquier R.A., Gobbi C., Wiendl H., Granziera C., Kappos L., Trendelenburg M., Leppert D., Lunemann J.D, & Kuhle J. (2024). Complement Activation Is Associated With Disease Severity in Multiple Sclerosis. Neurology(R) neuroimmunology & neuroinflammation, 11(2).

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Serum Biomarkers for Neurodegenerative Disorders

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Serum RPR and treponemal test reactivity, CSF WBC enumeration and CSF-VDRL reactivity were determined in a Clinical Laboratory Improvement Amendments (CLIA)-approved hospital laboratory. Plasma HIV RNA concentration, and peripheral blood CD4+ T lymphocyte concentration collected within 90 days of the LP were obtained from medical records. Serum NfL concentration was measured using the Simoa (single molecule array) NF-Light assay on an HD-X instrument according to instructions from the manufacturer (Quanterix, Billerica, MA). Serum was frozen at −20C and thawed just before analysis. All samples were analyzed in one round of experiments using one batch of reagents by board-certified laboratory technicians who were blinded to clinical data. Calibrators were run in duplicates. Samples were run in singlets with a 4-fold dilution. For a quality control (QC) sample with a concentration of 24.8 pg/mL, the intra-assay coefficient of variation (CV) was 5.5%. For a QC sample with a concentration of 73.7 pg/mL, the intra-assay CV was 7.1%.

Marra C.M., Sahi S.K., Tantalo L.C, & Zetterberg H. (2022). Serum Neurofilament Light in Neurosyphilis: A Pilot Study. Sexually transmitted diseases, 50(1), 42-44.

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Plasma NFL Concentration Comparison

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Plasma NFL concentration was measured using two highly correlated methods, employing the same antibodies: the in-house Simoa NFL assay that has been described in detail previously, 16 (link) and the commercially available NF-Light assay (Quanterix, Billerica, MA). Samples were analysed 'blind' and in duplicate using one batch of reagents. For the UK samples, an aliquot of the original baseline sample was analysed in the same batch as the 6-year follow up sample.

Rossor A.M., Kapoor M., Wellington H., Spaulding E., Sleigh J.N., Burgess R.W., Laura M., Zetterberg H., Bacha A., Wu X., Heslegrave A., Shy M.E, & Reilly M.M. (2022). A longitudinal and cross-sectional study of plasma neurofilament light chain concentration in Charcot-Marie-Tooth disease. Journal of the peripheral nervous system : JPNS, 27(1).

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