DENOVX, LLC — Department of Health and Human Services SBIR Phase II: 100

Description

PROJECT SUMMARY The goal of the Phase II grant is to expand the Phase I demonstration of improvements to protein crystallization achieved using engineered nucleation surfaces to other high impact targets that include proteins involved in antibiotic resistance and membrane proteinsThe role of crystallography in establishing the structure of biological macromolecules plays a key role in understanding their functionand Public Health benefits will derive from more efficient targeted drug developmentCrystallization of proteins remains a challengeand current approaches are experimentally intensive and have andgtfailure rateswhich creates acrystallization bottleneckthat is the rate limiting step in structural biologyPhase I Aims were accomplished using surface energy modifications giving an average reduction in crystallization onset times ofwith afold increase in the number of crystals experimentof whichformed on the engineered nucleation features for bovine pancreatic trypsinthaumatinlysozymeandlactoglobulinneachPhase II results with the first challenge protein PAalso known as NPprovided by the Center for Structural Genomics of Infectious Diseases at Argonneare even more remarkablethe engineered nucleation features improved crystallization hits byfold and produced afold increase in crystals inless timeDeNovX s products will be HTS plates for crystallizationand a slot in strategy will foster technology adoption by fitting into existing workflows and equipmentThe innovative aspects of the patented technologies are surface modifications that give tunable surface energies that enhance crystal nucleationOur hypothesis is that success outcomes for difficult to crystallize proteins can be improved using surface energy modifications to improve crystal nucleationSpecific AimExpand the ongoing probe of crystallization function to protein targets involved in the emerging threats from antibiotic resistancee glactamasesetcSpecific AimInitiate systematic investigations that advance the mechanistic understanding of how different surface energy modifications improve crystal nucleationThis understanding will pave the way for rational tailoring of the surface energy modifications for use with other soluble andmembrane proteinsSpecific AimExpand end user research and prototyping and initiatesite testing making HTS formats with surface energy nucleation enhancements available for use in membrane protein facilitiesThis equipment request seeks to fund acquisition of a dynamic light scatteringDLSplate reader that will permit the investigators to quantitatively probe protein aggregation in samples to be crystallizedProtein aggregation can inhibit or enhance crystal nucleationand a better understanding of protein aggregation size characteristics as a function of solution conditions and surface energy modifications will better equip the investigators to improve crystallization outcomes for high impact proteins and to diagnose the issues for those proteins that have failed prior screening attempts Project NarrativeRelevance to Public Health Research The role of structural biology in facilitating an understanding of biological function is well establishedand advances in crystallization methods will assist researchers in overcoming crystallization as a major source of attrition in their investigative pipelinesImprovements in protein crystallization outcomes will positively impact productivity in structural biologywith Public Health benefits accruing from faster knowledge acquisition and more efficient drug development