Array Imaging digital PCR Platform

Abstract Combinati is looking to bring an integrated Array Imaging digital PCR platform with injection molded microfluidic consumables from lab to marketplace to accelerate the adoption of dPCR technology. Digital PCR has drawn attentions in both genomic research and clinical research communities for its ability to detect rare events (high sensitivity), less prone to inhibition (high specificity), no need for an internal standard (high precision), and access to well-developed PCR reagents. However, current dPCR instruments are capital intensive, cost per sample is high, and the workflow is complicated. With successful completion of our Phase 1 aims, we will continue the following product development effort to bring our platform to market with the following differentiated features• Smooth transition from qPCR:o Equivalent workflow: The user will add the reagent mix to the consumable, reagentdigitization, thermal cycling, and real-time imaging is vertically integrated into one instrument.o Cost efficiency:▪ Instrument: The advance in imaging, thermal control and pneumatic controltechnologies allow us to sell the instrument at ~$30k, similar to qPCR instruments.▪ Consumable: The scalable injection molding technology enables low cost per datapoint (andlt;$1).o Open assay chemistry: The reagents will be partitioned in plastic microchambers insteadof proprietary emulsion oil. The user’s qPCR recipe can be directed transitioned to ourdPCR platform.o Automation compatible: The standard plate frame allows the user to automate the dPCRprocess with liquid handlers. • More robust and consistent data:o Real-time imaging during thermal cycling: By subtracting from background pre-thermalcycling image from the post-thermal cycling image, as well as recording amplificationdynamics of each partition, the user is able to count only partitions showing PCR tominimize false positive calls.o Fixed number of partitions for every data point: Poisson statistics is dependent on thetotal number of partitions. Our consumable will guarantee 20k partitions for each datapoint, unlike ddPCR, which varies between 11k to 19k.o Known volume: Quantification is done by dividing the number of molecules to the totalvolume. The volume of our array will be measured with confocal laser interferometry togive the user precise volume for quantification.Narrative There are increasing evidences showing that nucleic acids could complement proteins as additional biomarkers when it comes to disease prevention, prognosis, diagnosis, as well as therapeutics efficacy monitoring. Combinati is developing a precision nucleic acid quantification platform in the hope to accelerate the adoption of dPCR technology in the genomic research market, and catalyze its transition from benchtop to bedside. With the help from this Phase 2 SBIR project, we will deliver an integrated Array Imaging digital PCR (dPCR) platform from lab to marketplace with the best combination of workflow easiness, cost/sample, throughput/run and performance/experiment for high sensitivity absolute quantification of genetic biomarkers, targeting circulating cell-free DNA mutation detection as our beachhead application (recommended by our scientific collaborator ThermoFisher).