Thebiologicalfunctionandphysicochemicalpropertiesofprotein-basedtherapeuticsaredeterminedbytheir higherorderstructures(HOS).Therefore,itisimperativetoanalyzethethree-dimensionalhigherorder structureofproteintherapeuticsatseveralstagesofthedrugdevelopmentprocesstoensurebothsafetyand efficacyofthedrug.Biopharmaceuticalmanufacturersarerequiredtodemonstratetheconsistencyofthe proteinHOSconformationtotheregulatoryagencies.Moreover,Identifyingthebindingsiteofaproteintoits correspondingantigen(knownasepitopemapping)iscriticalforthedevelopmentofnewtherapeutics, vaccinesanddiagnostics.FoodandDrugAdministration(FDA)guidelinesrequirespecificbindingsite informationbetweenadruganditstargetfortheregulatoryfiling.CurrenttechniquesforproteinHOS characterizationandepitopemappingareslow,expensiveanddifficulttoperform.Wehavedevelopeda technologycalledPlasmaInducedModificationtoBiomoleculesthataddressestheneedoftheindustryfor routinestructural,massspectrometry-basedproteinHOSanalysis.PLIMBgeneratessubmicrosecondbursts ofhydroxyl(OH)radicalsfromwatertolabelproteinsinsolution.TheOHradicalscovalentlylabelsthesolvent accessibleregionsoftheproteinandsubsequentmassspectrometricanalysisrevealssingleaminoacidlevel structuralinformation.Awaytoquicklyandefficientlyanalyzehigherorderstructuresofproteinsonabenchtop scale,PLIMBwillenablefasterdevelopmentofproteintherapeutics,throughoutthedrugdiscoveryprocess.In SBIRPhaseI,wewillprovethefeasibilityofahydroxylradicaldetectionsystemthatwilltightlycontroltheOH radicaldosegeneratedbyPLIMBtoyieldhighlyreproducibleHOSdata.AftercompletionoftheSBIRPhaseI project,wewillcontinuethedevelopmentandvalidationofthePLIMBsystemforcommercialuse.Ultimately, PLIMBwillenablefast,highresolutionstructuralanalysisofproteins,acapabilitywhichishighlysoughtafterin thepharmaceuticalindustry.

Public Health Relevance

ThegoalofthisSBIRistodevelopahighlyreproducible,benchtopinstrumenttoperformHydroxylRadical ProteinFootprinting(HRF)foranalysisofthree-dimensionalstructuresinproteintherapeutics.Thisinstrument willenablestructuralcharacterizationofproteinsatamuchhigherthroughputthantraditionaltechniquesto betterdrivethediscoveryanddevelopmentofproteintherapeutics.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Small Business Innovation Research Grants (SBIR) - Phase I (R43)
Project #
1R43GM134849-01
Application #
9847255
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Krepkiy, Dmitriy
Project Start
2019-09-01
Project End
2020-08-31
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
1
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Immuto Scientific, Inc.
Department
Type
DUNS #
116833931
City
Madison
State
WI
Country
United States
Zip Code
53719