Fig. 1. Principle of the PEA-NGS method and description of cohorts.

a DNA-conjugated antibodies bind to their target protein if present in the sample. b Antibodies with correct matched pairs can hybridize and are extended by DNA polymerase. c DNA-tags on correctly matched antibodies are extended and amplified through PCR amplification. d Unique barcode-regions identify each sample and protein and adaptors allow for NGS amplification and read-out. The color code indicates different barcodes. e NGS generates a digital signal, which translates to the number of DNA hybridization events that corresponds to the protein concentration in the original sample. f Relative protein concentration levels are obtained after normalization and quality control. g The number of protein targets in PEA-NGS as compared to the previous method based on qPCR assay. h Schematic description of the two analyzed cohorts.