Project Summary The ultimate goal in the field of proteomics is single-molecule, high-accuracy, de novo protein sequencing. Unfortunately, current technologies used to characterize and identify proteins are costly, and lack the sensitivity, dynamic range, throughput, scale, and accuracy needed to meet the ever-expanding needs within this field. During this Phase I SBIR project, Electronic BioSciences, Inc. (EBS) aims to develop a nanopore reader, along with the associate methodology, capable of direct, single-molecule protein sequencing. A sensor capable of examining single proteins will not only provide detailed information regarding the individual protein themselves, it will also enable proteome-wide profiling and protein characterization, biomarker(s) discovery, and the associated assessments of disease state progression or remission. The prototype sensor developed here will be a high-accuracy nanopore-based reader capable of label-free sequence characterization that provides high- quality data without sequence-context convolution. This development will be the first of its kind, ideally suited to assessing and sequencing any given protein, regardless of length and structure. Additionally, the sensitivity of the proposed reader will enable the ability to detect and identify specific post translational modifications, which is critical to understanding and assessing the extensive variation within the proteome.