Direct Proteolytic Machine Assessments for Unfolding and Sequencing Proteins

Project Summary Protein sequencing is a cornerstone of proteomics, offering crucial insights into the molecular underpinnings of biological processes and diseases. However, current proteomic tools, such as mass spectrometry (MS), immunoassays, and protein cataloging approaches, are limited by high costs, low throughput, and an inability to perform comprehensive de novo protein sequencing. These limitations hinder the accurate characterization of low-abundance proteins and the sequencing and quantification of post-translational modifications (PTMs), single nucleotide polymorphisms (SNPs), and splicing variants, making proteome-wide analysis challenging. Given these constraints and the growing demand for more comprehensive proteomic analysis, there is a significant opportunity to develop more efficient, cost-effective, and scalable protein sequencing methods. In response, Electronic BioSciences, Inc. (EBS) proposes to develop and demonstrate a novel enzyme-based, single- molecule protein sequencing system. This innovative platform will leverage enzyme-driven processing and sequencing of individual proteins and peptides, combined with advanced bioinformatics, to achieve high accuracy and throughput. During this Phase I program, we will develop and optimize this single-molecule sequencing approach, demonstrating its ability to accurately sequence peptides with various PTMs, surpassing the capabilities of current state-of-the-art methods. By integrating enzymatic assessments with sophisticated sequencing algorithms, our system aims to enable scalable, multiplexed protein sequencing, capable of analyzing all proteins and peptides in a given sample. Our proposed system will not only be rapid, reliable, and low-cost but also accessible for a wide range of applications, from basic research to clinical diagnostics. This technology has the potential to revolutionize protein analysis, facilitating rapid biomarker discovery and enhancing disease testing and monitoring, ultimately advancing biomedical research and public health.