Msd 5 plex human cytokine 30 plex kit

EDTA-plasma samples were analysed using the MSD V-PLEX Human Cytokine 30-plex kit (K15054D; Mesoscale Discovery, Gaithersburg, Maryland, USA) according to manufacturer’s instruction. In short, the plasma samples were thawed at room temperature and just before analysis diluted, twice for proinflammatory cytokine and four times for chemokine analysis, in sample diluents. The diluted samples were incubated on the MSD plates for 2 hours at room temperature while shaking. Plates were washed and incubated in additional 2 hours with detection antibodies. After washing, 2× Read buffer T was added to each well and the plates were analysed in a Sector Imager 6000. Lower limit of quantification (LLOQ) was determined for each assay plate based on the lowest standard point with a coefficient of variation  <25% and an accuracy (back-calculated concentrations) of 80%–120%.

Cytokines were analysed in EDTA plasma samples from the SLE cohort using the MSD V-PLEX Human Cytokine 30-plex kit (K15054D; Mesoscale Discovery, Gaithersburg, Maryland, USA)33 and cytokines, which previously have been suggested to be increased in patients with SLE,34–37 (link) that is, tumour necrosis factor alpha (TNF-α), IFN-γ-induced protein 10 (IP-10), interleukin (IL)-16, macrophage inflammatory proteins 1 beta (MIP-1β), and IL-6 were studied in this work.