Generating sandwich assays through MegaSTAR

Project Summary The andapos immunosandwichandaposassay is a robustflexible and sensitive test that is widely employed in clinical diagnostics for monitoring human health and diseaseIt is based on using pairs of antibodies that recognize different epitopes of the target analytethereby providing both sensitivity and specificityUnfortunatelyidentifying pairs of antibodies that work in sandwich assays is problematici etakes time and luckparticularly when they are obtained from animal sourceswhere there is no specific selection for complementarityThe development cost of a sandwich assay approximates $ MThis technology gap can be filled with recombinant affinity reagentswhich are sequencedrenewableand amenable to protein engineeringTango BiosciencesInchas licensed intellectual property for isolating complementary pairs of affinity reagentsas part of the phage display affinity selection processrather than iterative testing post productionThis recently published technologynamed MegaSTARyieldstodifferent pairs of affinity reagents for targets in one monthandapos s timeThe STTR award will assist in transitioning the technology from an academic center to a commercial entity that can develop a standard operating protocolActualization of the company will result from the successful completion of three specific aimsIn Specific Aimwe will prepare two sets of targets for affinity selectionOne setcarcinoembryonic antigenCEAbiomarker of colorectal cancerand cardiac troponin TcTnTbiomarker of myocardial infarctionwill serve to benchmark our reagents against commercial kits that utilize polyclonal and monoclonal antibodiesThe second setleukocyte immunoglobulin like receptorsLILRB typepotential immune checkpoint proteinswill serve as challenging new targets for which monospecific antibodies are not currently available to their ectodomainsIn Specific Aimwe will use MegaSTAR to generate and optimize pairs of affinity reagents for each targetIn Specific Aimwe will compare our reagents and commercially available kits for detecting CEA and cTnT in spiked human plasmaThe performance metricsi eintra assay and inter assay precisionlinearitysensitivityof reagents will be evaluated through ELISASeparatelywe will evaluate our affinity reagents to the five LILRB ectodomains for their relative specificity through ELISASuccessful completion of the proposed experiments in themonth period will enable Tango Biosciences to demonstrate the commercial utility of MegaSTAR and solicit research service contracts from reagent companies and applied marketsThe realization of Tango Biosciencestechnology platform will positively impact research and clinical testing in ways that potentially improve health treatment and outcomes in the US Project Narrative Human health and disease can often be monitored through biochemical testsThis NIH STTR Phase I project will support an effort to develop and apply an innovative protein engineering technology into commercial products that enable researchers to detect disease and infection on demand