Digital Microfluidic Artificial Golgi for Glycan Synthesis

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Digital Microfluidic Artificial Golgi for Glycan SynthesisJonathan DordickRensselaer Polytechnic InstituteCBET-0730817Glycosylation, the most important posttranslational modification of proteins, occurs in the Golgi organelle. The complexity of glycosylation has hindered fundamental understanding of biosynthesis and has made controlled synthesis of essential therapeutic agents difficult and expensive. Therefore, there is a compelling need to develop a fundamental understanding of glycan biosynthesis within the Golgi, which will lead to the development of artificial routes to the synthesis, modification, and ultimately large-scale synthesis of glycans and glycoproteins. The goal of this research is to perform the controlled biocatalytic synthesis of heparan sulfate/heparin via digital microfluidic systems (DMFS), which are a new class of lab-on-chip systems that manipulate discrete droplets of chemicals on an array of electrodes. Intellectual Merit: This research will lead to a fundamental understanding of glycosylation in the Golgi, and mimic these processes by examining precisely controlled variations of spatial and temporal parameters in enzyme catalysis. As a model, in-vitro synthesis of heparan sulfate/heparin will represent the first demonstration of an artificial Golgi that may be of utility in the synthesis of a wide variety of therapeutic glycoproteins. Precise control algorithms will be developed to enable efficient glycosylation and will establish a comprehensive framework to design fluid movement, retention, and separation, much like in the Golgi organelle. Broader Impacts: Results will be integrated into graduate courses on Synthetic Biology and Robot Motion Planning. An even broader impact will result from integrating this project into ongoing efforts at RPI in the development of the MoleculariumTM, a combination planetarium and virtual space ride that takes the audience into the world of atoms and molecules. Broader societal outcomes of the proposed research include the development of the next generation of glycan-containing drugs and the design of low-cost, portable lab-on-a-chip systems capable of rapid automated biochemical analyses for point-of-care testing, research and drug discovery.