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