The article presents a novel aptamer-based proteomic technology for biomarker discovery, capable of measuring thousands of proteins from small sample volumes (15 μL of serum or plasma). The technology uses a new generation of aptamers that contain chemically modified nucleotides, expanding the physicochemical diversity of the libraries. The assay measures 813 proteins with low limits of detection (1 pM median), a 7-log overall dynamic range (~100 fM–1 μM), and a 5% median coefficient of variation. The technology is demonstrated through a clinical study of chronic kidney disease (CKD), where it identified two well-known CKD biomarkers and 58 additional potential biomarkers. The results show the potential of this technology to rapidly discover unique protein signatures characteristic of various disease states, advancing the next generation of evidence-based medicine.The article presents a novel aptamer-based proteomic technology for biomarker discovery, capable of measuring thousands of proteins from small sample volumes (15 μL of serum or plasma). The technology uses a new generation of aptamers that contain chemically modified nucleotides, expanding the physicochemical diversity of the libraries. The assay measures 813 proteins with low limits of detection (1 pM median), a 7-log overall dynamic range (~100 fM–1 μM), and a 5% median coefficient of variation. The technology is demonstrated through a clinical study of chronic kidney disease (CKD), where it identified two well-known CKD biomarkers and 58 additional potential biomarkers. The results show the potential of this technology to rapidly discover unique protein signatures characteristic of various disease states, advancing the next generation of evidence-based medicine.