Sample Collection and Measurement of Serum Neurofilament Light (NfL)

Abstract

This protocol details the steps for the preparation of serum from human blood samples and the measurement of NfL concentration using the NF-Light Advantage kit on the HD-X Analyzer (Quanterix, Billerica, MA).

Before start

Check the planned procedure is covered by existing Research Ethics Committee approval and other required approvals.

Check that you have the correct personal protective equipment, supplies and consumables.

Steps

Blood Sample Collection and Serum Preparation

At enrolment, collect 10 mL of venous blood from each participant in Serum Separator Tubes (SST): https://www.fishersci.co.uk/shop/products/vacutainer-sst-ii-advance-tubes/12927696

Safety information

Venepuncture and the handling of blood has the potential to expose research andclinical personnel to blood borne pathogens. In order to minimise risk, personnelperforming a venepuncture must have received appropriate training and deemedcompetent. Institutional health & safety policies, risk assessments and standardoperating procedures must be followed.

Within 1 hour of collection, centrifuge the SSTs at 2,500g for 15 minutes at room temperature

Print cryogenic labels / barcodes and apply to cryotubes: https://www.brady.co.uk/labels

Aliquot approx. -0.5mL serum into labelled cryotubes, box and/or rack then store at -80°C.

Measurement of Serum NfL Concentration

Measure serum NfL concentration using the Simoa® NF-light Advantage Kit for HD-1 / HD-X following the manufacturers instructions : https://www.quanterix.com/wp-content/uploads/2020/12/NF-light-Data-Sheet-HD-1%E2%88%95HD-X-2.pdf

This kit is a digital immunoassay for the quantitative determination of NfL in serum, plasma and CSF.

Note

*The measurement of serum NfL concentration is performed blinded to clinical diagnosis.

Run on the HD-X Analyzer (Quanterix, Billerica, MA), a fully automated Simoa® bead-based immunoassay platform: https://www.quanterix.com/instruments/simoa-hd-x-analyzer/