Fig. 3: Performance of AEMS for 10-plex APR assay.

A The dynamic range of the 10-plex APR was explored in pooled human plasma using a standard addition curve, spanning a range of almost > 1000000, highlighting their varying abundance levels and potential implications for physiological processes. B Reproducibility of endogenous peptide areas for the acute phase response peptide areas from plasma captures (n = 268). As expected, the reproducibility observed for the endogenous peptides from triplicate technical measurements correlates with the observed peptide area or abundance, with the higher abundance peptides showing very good reproducibility (<10%) and the less abundant, lower area peptides having more variance. Data was subjected to the outlier rejection strategy and rejected data points were not plotted. Inset shows the reproducibility of acute phase response SIL peptide areas from plasma captures, in a violin plot. Data are represented by the median, first and third quartiles, and range. The reproducibility of the SIL peptide for each enriched sample measured in triplicate was found to be very good, with average %CV values across the 267 measured samples between 4.2% and 10.5%. C Correlation of measured AEMS L/H peptide ratios with LC-MS data (n = 71) for the most biologically relevant acute phase response proteins, namely A1AG, C3, LBP, CRP and SAA. The ratios measured by LC-MS were very similar to the ratios determined using the Echo MS system. After outlier rejection, the slopes for all proteins were very close to 1 and the R² values were 0.96 and higher.