Fig. 3: Characterization of nELISA specificity, sensitivity and reproducibility.
a, A schematic representation of rCR in traditional multiplexed ELISAs and its abrogation by CLAMP. b, Correlation of signal-to-noise ratio (SNR) values for CLAMP(s) in 1-plex or 191-plex format; an example standard curve is also shown (inset). c, Spike-one-in assay: heatmap displays SNR values for each nELISA sensor in the 191-plex for each individually spiked antigen; diagonal displays specific signals; cross-reactive events are numbered. d, Leave-some-out assay: six pools of recombinant proteins consisting of either all the targets in the 191-plex panel (A1) or lacking a subset of targets (A2–A5) were profiled; outlined boxes indicate absence of target proteins; colors represent SNR. e, Overlaid standard curves for one example CLAMP (CCL5) using (red) standard and (blue) xDR protocols. f, Distribution of SNR values for all 191-plex across 80 cell culture supernatants from stimulated PBMCs quantified using (red) standard or (blue) xDR protocols. g, Distribution of the lower limits of detection (LLOD) and upper limits of detection (ULOD) of 191 sensors using standard or xDR protocols. h, Distribution of coefficients of variation (CV) of all nELISA sensors in repeat measurements of a single sample across wells in a single plate (well to well) and across plates profiled on different days (day to day). i, Cell culture supernatants from stimulated PBMCs were analyzed with nELISA and xMAP platforms; shown is the distribution of Spearman correlation coefficients for shared assays with detectable protein concentrations and an example of correlating IL-1β concentrations (inset). j, Cross-reactivity comparison on xMAP and nELISA platforms: 100 recombinant antigens were pooled and spiked in cell culture media at increasing concentrations; shown are the quantification of seven proteins that are not detectable in the sample using the nELISA (top) and xMAP (bottom).
