Plasma research analysis
Plasma samples collected immediately before and 1, 3, 6, 12 and 24 months after transplant from the 12 treated patients were analyzed for NF-L, GFAP, Tau and UCHL-1 content using Simoa Human Neurology 4-Plex A (N4PA, 102153) digital immunoassay on an HD-1 single-molecule array (Simoa) instrument according to instructions from the manufacturer (Quanterix). The measurements were performed by a technician blinded to clinical data, in one round of experiment using one batch of reagents and with baseline and follow-up samples analyzed side-by-side. Plasma samples were diluted four times. This is a two-step immunoassay, in which target antibody coated paramagnetic beads are combined with sample and biotinylated detector antibody in the same incubation. Target molecules present in the sample are captured by the antibody-coated beads and bind with the biotinylated antibody detector simultaneously. After a wash, a conjugate of streptavidin-ß-galactosidase (SBG) is mixed with the beads. SBG binds to the biotinylated detector antibodies, resulting in enzyme labeling of the captured target. After a final wash, the beads are resuspended in a resorufin ß-