Quantification of Neurofilament Light by Simoa

For NfL measures, samples were thawed for 60 min at room temperature and were analysed by an investigator blinded to clinical data using the Simoa Nf-light kit in the Simoa HD-1 Analyser (Quanterix, Lexington, MA, USA), which runs ultrasensitive paramagnetic bead-based enzyme-linked immunosorbent assays [27 (link)]. For this protocol, briefly, thawed samples and calibrators were dispensed in provided 96-well plates as duplicates. Further sample processing (dilution, incubation, washing, shaking, resuspending and reading) was carried out in an automated manner as described elsewhere [28 (link)]. The sNfL assay was carried out according to the manufacturer’s instructions and protocol.