Unmet Need
Protein cleavage, catalyzed by enzymes (proteases) that break peptide bonds, is a fundamental molecular process essential to cellular viability and tissue physiology across all kingdoms of life. Protein cleavage plays pivotal roles in tissue remodeling and morphogenesis, infection, blood coagulation, neoplasia, and cancer metastasis. The enzymatic activities of proteases can serve as valuable diagnostic biomarkers, therapeutic targets, and a method to monitor a broad range of biological functions, yet assays to characterize these enzymatic activities at a proteomic scale do not exist. There is a need for a comprehensive assay to analyze and characterize the entire set of proteases of a biological sample to understand their structure, function, interactions, and how they change under different conditions and disease states.
Technology Overview
JHU’s novel technology leverages a phage display-based platform to provide unparalleled insights into protease activity. The system is designed to meet the critical need for comprehensive assays that characterize protease activities within biological samples, offering significant advantages for research and clinical applications. JHU researchers developed Sensing EndoPeptidase Activity via Release and recapture using flAnking Tag Epitopes (SEPARATE), a highly multiplexed protease-profiling platform that combines a complete human proteome library cloned into a custom designed T7 phage display vector and quantitative analysis via next-generation DNA sequencing. SEPARATE enables unbiased, low-cost, and high sample throughput characterization of human protease activities, thereby overcoming key limitations of current approaches.
Key Benefits:
- High Sensitivity: Utilizing DNA analysis for detection, our platform achieves extremely high sensitivity, ensuring accurate and reliable results.
- Comprehensive Profiling: The system encodes complete proteomes, allowing for the unbiased, quantitative detection of all peptide cleavage events.
- Versatility: Applicable across various fields, including oncology, immuno-oncology, and in vitro diagnostics, making it a versatile tool for diverse research needs.
- Efficiency: The phage display system requires minimal time and resource expenditure, while enhancing throughput.
Stage of Development
Prototypes have been completed and are in the testing phase.
Patent Information
PROTEASE ACTIVITY PROFILING VIA PROGRAMMABLE PHAGE DISPLAY OF COMPREHENSIVE PROTEOME-SCALE PEPTIDE LIBRARIES Pending United States application: 16/913,572
Filed: 6/26/2020
https://patents.google.com/patent/US20200407467A1/
Publication
Gabriel D. Román-Meléndez, Thiagarajan Venkataraman, Daniel R. Monaco, H. Benjamin Larman,
Protease Activity Profiling via Programmable Phage Display of Comprehensive Proteome-Scale Peptide Libraries,
Cell Systems, Volume 11, Issue 4, 2020, Pages 375-381.e4, ISSN 2405-4712
https://doi.org/10.1016/j.cels.2020.08.013.
(https://www.sciencedirect.com/science/article/pii/S2405471220303240)
