The Bilayer Nanopore Integrated Assay for Complete Toxin Characterization, Phase II

Project Summary During this programElectronic BioSciencesEBSwill further developvalidateand begin commercializing its in vitrocartridge basedbilayer nanopore integrated assayBNIAfor characterizing the complete activity of transmembrane toxinsA transmembrane toxin is a molecule that recognizes targets a specific cell type via receptor mediated targeting endocytosisforms a pore in the cell membraneand transports itself or another molecule into the cell to disrupt cellular functione gbotulinum neurotoxinBoNTtetanusTeNTdiphtheriashigacholerapertussisetcWhile these toxins are innately hazardous to human healththeir inherent cell targeting and enzymatic capabilities can also be harnessed for therapeutic benefitTodaythe emerging therapeutic uses of toxins include but are not limited to the treatment of muscle spasmswrinklesexcessive sweatingdepressionanxietyanorexianeurodegenerative disorderse gParkinson s diseaseand targeted cancer therapyHoweverthe limitations of current toxin activity assessment methods have constrained the fieldTodaythere is a need for a low costeasy to userapidhighly sensitivehighly reproducible assay that is capable of individually quantifying the separate steps of the intoxication mechanismi ethe cell targeting endocytosis and the intracellular enzymatic activityto fully understand and utilize toxin functionalityThe present standard for toxin activity assessments is the mouse intraperitoneal injection assaywhich has numerous limitationsincluding pricevariabilitytimelack of sample quantificationand the utilization of live animalsin addition relying on a single endpoint determination that precludes assessment of the toxin s mechanismUnknown or poorly understood differences in the potencyor mode of actionof toxin containing therapeutics can confound clinical dose findingsresult in over or under dosing patientsand delayor prohibitthe development and or availability of new novel therapeuticsEBSBNIA technology is capable of unprecedented toxin characterization in a low costeasy to userapidhighly sensitivehighly reproduciblein vitrocartridge based platformFurthermorethe methodology of the BNIA is customizable such that the complete activity of any transmembrane toxin can be quantifiedThe further development and commercialization of the BNIA under this Phaseprogram will be accomplished by further demonstrating the BNIA s utility and enabling featuresdeveloping BNIA beta prototypesand introducing the developed BNIA to the academic and industrial user baseThe developed BNIAa technology for which there is no equivalent commercially available and the future gold standard in toxinbiotherapeuticscell targetinguptake internal activityand causation mechanism quantificationwill directly enable the research and development of novel toxin agents samplesthe development of antitoxin agentsthe detailed study of toxin and antitoxin mechanismsthe evaluation of the causative effects of experimental variables on each specific intoxication modalityand the assessment of toxin potency in general