Development of Engineered Trypsin for Proteomics Sample Prep.

ABSTRACT SignificanceNext generation sequencingNGShas has transformed the field of genomics and enabled broad adoption of the technology in biologicaltranslationaland most recently clinical researchThe proteomics equivalent to NGS has not yet been realizedAs suchmore accessible and cost effective technologies for analysis of the proteome would accelerate biological and medical researchThe goal of this proposal is to develop a highly scalable digital protein analysis technologyThe approach is broadly applicable and greatly improves upon costsensitivityusabilityand throughput of current approachesApproachWe are developing a single molecule digital peptide sequencing platform using highly parallelized single molecule Edman degradation and DNA encodingOur assay generates a DNA library representing peptide sequence information on millions to billions of peptides that is sequenced on a standard NGS instrumentIn this proposalwe are focused on developing reagents to expedite DNA tagging of the millions to billions of peptides for our assay by engineering proteases to cleave at modified sitesImpactIn summarywe believe our Next Gen peptide sequencing assay will usher in the era ofnext gen proteomicseffectively transforming biologicaltranslationaland clinical researchmuch like NGS technology has already done in the genomics space NARRATIVE This proposal is to develop reagents technology to enable user friendlyhigh throughput analysis of proteinsBy making information about proteins more easily accessiblethis technology has the potential to advance basic research in almost every area in molecular life sciencesas well as applied and clinical researchincluding drug target discovery and drug development