Methodology

Saliva is tested on the V-plex pro-inflammatory cytokine panel (Ref# K008074) using the Meso Scale Discovery (MSD)® multi-spot Assay system (MSD® www.mesocale.com). In this system, an array of capture antibodies against the different targets of interest is patterned on distinct spots in each well of a solid phase 96-well electrode surface plate. Standards and unknown samples are added to the plate and incubated while shaking at room temperature for 2 hours. The plate is washed and then a master mix containing detection antibodies (Abs) labeled with an electrochemiluminescent compound (MSD SULFO-TAG™) is added, and the plate undergoes another 2 hour shaking incubation. As in a traditional sandwich ELISA, analytes in the sample bind to the immobilized, arrayed, capture antibodies on the working electrode surface and recruitment of the labeled detection Abs by captured Abs completes the sandwich. A read buffer is added to provide the chemical environment for electrochemiluminescence and the plate is read in a MESO QUICKPLEX SQ 120 (MSD®) which applies a voltage to the plate electrodes causing the bound detection labels to emit light. The instrument measures the intensity of emitted light.