CATASSAYS — Department of Health and Human Services SBIR Phase I: NCI

Description

DESCRIPTION provided by applicant We propose to use CatAssaysandapos novel Palladium Catalyzed Chemical Amplification PdCCA technology platform to develop ultra sensitive sandwich ELISA format bioassays to detect very low concentrations of a number of critical cancer and immunoregulatory biomarkers CatAssaysandapos PdCCA technology the first example of the application of heterogeneous metal catalysis in bioassays improves the signal generation chemistry while using nd party highly specific state of the art capture detection antibodies t provide a significant increase in system detection sensitivity by a use of heterogeneous palladium nanoparticle catalyst labels in place of an enzyme catalyst used in conventional ELISA b use of a new high gain redox reaction the palladium catalyzed reduction of a tetrazolium salt by an amine borane to give a water soluble formazan dye Initial reduction to practice of PdCCA in which the detection antibody itself was labeled with palladium gave an fold detection sensitivity increase vs the standard enzyme based ELISA test for t PSA Gen PdCCA The proposed work will develop a more commercially viable nd generation PdCCA technology based on the use of a universal detection label comprising polymeric nanoparticles containing a high loading of the palladium catalyst and surface functionalized with a biospecific binding reagent e g Steptavidin etc for attachment to any biotinylated detection antibody This Phase work will focus on synthesis of the above universal catalytic polymeric marking nanoparticles and optimization of Gen PdCCA technology in bioassays using model biomarkers initially TNF IFN IL followed by IL IL IL cytokines and CA SLPI and MSLN ovarian and tPSA cancer biomarkers to demonstrate improved sensitivity for a broad range of applications The implementation of optimized PdCCA technology incorporating the new palladized polymeric marking nanoparticles in modified ELISA format assays is not time consuming making the Phase evaluation of the above biomarkers readily achievable In Phase a viable commercial scale synthesis for the Gen PdCCA catalytic polymeric marking nanoparticles and assay kits for specific cancer biomarker assays will be developed At least one ovarian cancer biomarker PdCCA assay identified in collaboration with our medical center experts will be submitted for FDA approval This new bioassay signal amplification technology is expected to provide a readily implementable and cost effective significant increase in sensitivity with no detectable loss of specificity for the earlier detection of cancers while using the current medical laboratory infrastructure PUBLIC HEALTH RELEVANCE CatAssaysandapos proprietary ultrasensitive high gain amplification technology Fig used in this SBIR Phase proposal as a generic analytical platform can provide a significant increase in detection sensitivity for a broad range of target analytes and assay formats This technology addresses the critical need of the oncology community for increased detection sensitivity for highly selective antibodies that have been identified for a broad range of cancer biomarkers found in blood serum and other body fluids It will be especially useful where a critical need exists for increased sensitivity to provide earlier detectin of diseases allowing earlier treatment and decreased fatality rates as well as decreased cost of medical care associated with treatment of advanced stages of such diseases This technology is applied here as a modified sandwich ELISA enzyme linked immunosorbent assay assay the most widely used bioassay format which has the following critical features specificity the ability of chemically labeled antibodies to bind to target disease biomarkers with very high degrees of specificity in the presence of the large number of proteins in a serum sample signal generation the ability to provide a readable signal using a chemical reaction that is initiated by the label on the detection antibody In the standard ELISA bioassay an enzyme label is used as a catalyst for the hydrogen peroxide oxidation of a substrate to a dye signal CatAssaysandapos technology the first example of the application of heterogeneous metal catalysis in bioassays improves the signal generation chemistry while using nd party state of the art capture detection antibodies to provide a significant increase in system detection sensitivity by a use of heterogeneous palladium nanoparticle catalyst in place of an enzyme catalyst used in conventional ELISA b use of a new high gain redox reaction the palladium catalyzed reduction of a tetrazolium salt by an amine borane to give a water soluble formazan dye Palladium Catalyzed Chemical Amplification PdCCA In addition to enhanced sensitivity the other key feature of this technology is its compatibility with the standard ELISA format assay widely used in the medical diagnostic market without the need for any capital laboratory investment for implementation i e its implementation requires changes in the above chemical steps but the dye signal is detected using the same optical spectrophotometry for dye signal quantification as in standard ELISA technology Initial reduction to practice of PdCCA in which the detection antibody itself was labeled with palladium gave an fold detection sensitivity increase vs the standard enzyme based ELISA test for t PSA Gen PdCCA a The proposed work will develop a more commercially viable nd generation PdCCA technology based on the use of a universal detection label comprising polymeric nanoparticles containing a high loading of the palladium catalyst and surface functionalized with a biospecific binding reagent e g Steptavidin etc for attachment to any biotinylated detection antibody This Phase work will focus on synthesis of the above universal catalytic polymeric marking nanoparticles and optimization of Gen PdCCA technology in bioassays using model biomarkers initially TNF IFN IL followed by IL IL IL cytokines and CA SLPI and MSLN ovarian and tPSA cancer biomarkers to demonstrate improved sensitivity for a broad range of applications In Phase a viable commercial scale synthesis for the Gen PdCCA catalytic polymeric marking nanoparticles and assay kits for specific cancer biomarker assays will be developed