Overview of affinity proteomics screening and study design.

(A) Schematic overview of the affinity proteomics approach using antibody suspension bead arrays. Plasma samples were biotinylated, antibodies were coupled to color-coded magnetic beads, and both were combined for analysis. Bead identity and captured plasma proteins were then detected using a flow cytometric analyzer. (B) Experimental design of study. Initial screening with 1,132 antibodies from targeted and blinded selections was performed in the discovery cohort (n = 356). Data from the patient groups were compared using univariate tests and multivariate penalized regression models. Identified single proteins and multi-component protein panels discriminating the 3 disease groups were validated in the verification cohort (n = 363).