Cell-free protein synthesis (CFPS) devices with high-yield for synthetic biology

Project SummaryCell free protein synthesisCFPSis a method of producing proteins outside living cells and has distinct advantages over traditional cell culture based protein productionIt is often difficult to maintain complex environmental condition for cell viability and the subsequent need to develop a downstream process for purification of product in traditional cell based systemsCFPSon the other handcan be accomplished just by mixing genetic templateDNAor mRNAwith a reaction solution containing transcriptiontranslation componentse gribosomesand nutrient molecules in an in vitro setupThe synthesis time is also less than a day compared to weeks for the cell based systemMoreoverlack of cell membrane and open nature of CFPS system can be advantageously used for adding different molecules to optimize the protein synthesisConsequentlyCFPS is used for industrial scale therapeutic protein productionenzyme engineeringprotein labeling and studying protein protein interactionWhile the CFPS system has been around for a few decades and has high potential to transform the biosynthesis marketit still has some important issues like high reagent costlow protein yieldprotein aggregation and lack of post translational modification that has throttled its growthIn this projectwe will develop a universal CFPS device that will be able to synthesize proteins at a higher yieldlow cost of reagents and even less time to synthesizehours compared tohoursDasfanh Bioscience will use patent pending technology to solve the problem by introducing a membrane between reaction and feeding solutionand continuously supplying the nutrients to the reaction chamber and removing by products from the reaction chamberThis results in significant increase in the protein synthesis yieldone to two orders of magnitudewith a reduction in cost by at least one order of magnitudeSpecific AimsTo develop awell based microfluidic device that can be universally used for all forms of cell free protein synthesis with significantly higher yield in less timeTaskDevelopment ofwell based continuous exchange CFPS device for high throughput applications MilestoneDesign and develop the flexible membrane devices for maximum exchange of molecules aided by active transport of fluidMilestoneExtensive testing of commercially available CFPS reagentsboth prokaryotic and eukaryoticto validate efficiency and compatibility with the deviceTaskDemonstration of versatile usability of the system by co expressing various proteins and studying interactions among themTaskAdaptation of this technology for co generation of ribosomal proteins for synthetic biologyProject NarrativeCell free protein synthesisCFPScan provide a valuable alternative to traditional cell culture based platforms for protein productionThe new device to be developed in this STTR can be used with all types of CFPS systems with advantages in the synthesis yieldreaction efficiencyand throughputThis device will be compatible with other laboratory instruments like microplate readersincubatorsand robotic fluid handling equipmentThe expected customers will be pharmaceutical companies for drug discovery and screening studiesbiotechnology companies for small scale protein productionand research laboratories for protein protein interactionfunctional and crystallographic studiesand synthetic biology