Virus Size Classification Surveys to Monitor Superinfections

Project Abstract Electrospray Ionization Mass SpectrometryESI MSis a biochemical analysis workhorsewith Life Sciences equipment sales$billion annuallysee BibliographyIt is well developed for the analysis of proteinspeptidesDNA and macro moleculeswith sizes belownmbut is not yet able to cover the next register in sizesthe virus rangenmSchemeMobility based size analysis of charge reduced electrosprayed particles has offered an alternative window to the viral size range sinceHoweverin spite of outstanding demonstrated sensitivitysingle particle countand quantification abilityfar superior to other first line nonbiological tools such as electron microscopythe limited resolving power of its commercial formTSI Inchas thwarted its widespread adoptionThis SBIR promises to fulfill critical unmet need for quickgeneral purposevirus particle size analysiswith greatly improved resolving powerresulting in sharp distinction of NanoscopicVirionand large protein complexparticles in the rangenmHigh size resolution not only better distinguishes viral analytes for species identificationbut also makes viral peaks emerge distinctly from the backgroundespecially because in insect vector Superinfections one dominant viral contagion population suppresses all others through viral interferenceWe anticipate a size resolving power in excess ofcomparable with the outstanding recent developments in virus MS by Jarroldthough in a far simplereconomicalfaster and more sensitive deviceNanoEngineering Corporation sNECcurrent portable precision High ResolutionIonMobility Analysis of ESI aerosolized virus particlesNanoRangerinstrument offers already a resolution andgtover insect virus rangenmwith potential extension tonm to span nearly all respiratory virusesThis resolution well exceeds limits of conventional laser particle scatteringWe propose three main developmentsiExploit systematically the existing high resolution Differential Mobility AnalyzerDMAin collaboration between NECYale and BVS Incto develop improved protocols for sample preparationespecially of insect viriThis collaboration should be most fruitfulas no high resolution mobility instrument has been previously available to guide improved sample preparationiiFurther develop this DMA to increase its resolving powerThe improved DMA will be tested at Yale with Bio Safety Levelviral particles having a fixed sizei ea capsidiiiA final vital proposed development is to drastically advance the Tech Readiness Level of the existing size analysis DMA instrumentationsuch that it can reliably be used by virologists during Phase II of this programResults will qualify this SBIR tech for Phase II IIB trials for field surveillance of mosquitos borne contagionsZikaDenguePhase II will be accomplished via installation of a beta unit at Scripps Research Institute s BLSfacilityFLa stepping stone to deployment with early adopters at three mosquito labsUof SFLUSDAFL Medical Entomology Lab and beyondDMA Phase II extension will span tonmPhase III will target commercial manufacturing and marketing of patented NanoRangerESI DMA instrumentsESI Mass Spectrometrynm diameter MacromoleculesUnderserved Size Gapnm Virus Size RangetonmOptical Microscopesnm diameter Single Cell Organisms etcParticle SizenmSchemeUnmet market need for quicklow cost virion nanoparticle size measurement may be fulfilled by ESI DMASBIR success could serve as springboard to general adoption of this instrument in life sciences and biomedicine Project Narrative Existing mobility based size analysis of electrosprayed charge reduced viral particles will be developed to achieve much higher size resolution than previously possibleThe new instrument will be systematically used in collaboration with virologists to greatly improve virus sample preparation protocolsnarrowing viral size distribution to achieve high genericnon biologically distinctivevirus recognition capabilityThe reliability of the instrumentation will be greatly increased to enable its straightforward use in Phase III by three leading Laboratories to monitor mosquito populations for the prevalence of tropical diseasesIntroduction to Resubmission and Revision Applications The present proposal is a revision of NanoEngineering s ApplicationRGMFollowing referee s requestsit provides considerably more technical detail on the novelty of the proposed approachthe insect sample preparation protocoland the drastic advantages over existing low resolution mobility size instrumentation commercialized by TSI