Digital Microfluidic Artificial Golgi for Glycan Synthesis

用于多糖合成的数字微流控人工高尔基体Jonathan DordickRensselaer理工学院CBET-0730817糖基化是蛋白质最重要的翻译后修饰，发生在高尔基体细胞器中。糖基化的复杂性阻碍了对生物合成的基本了解，并使基本治疗药物的受控合成变得困难和昂贵。因此，迫切需要对高尔基体内的多糖生物合成有一个基本的了解，这将导致人工路线的发展，以合成，修饰，并最终大规模合成多糖和糖蛋白。本研究的目标是通过数字微流控系统(DMFS)进行硫酸肝素/肝素的受控生物催化合成，这是一种新型的芯片上实验室系统，可以在一系列电极上操纵离散的化学液滴。智力价值：这项研究将导致对高尔基体糖基化的基本了解，并通过检测酶催化中精确控制的空间和时间参数的变化来模拟这些过程。作为一个模型，体外合成硫酸乙酰肝素/肝素将代表着人工高尔基体的首次展示，这种人工高尔基体可能在合成各种治疗性糖蛋白方面有用。将开发精确的控制算法以实现高效的糖基化，并将建立一个全面的框架来设计流体的运动、保留和分离，就像在高尔基细胞器中一样。更广泛的影响：成果将被整合到合成生物学和机器人运动规划的研究生课程中。将该项目整合到RPI正在进行的MoleculariumTM开发工作中，将产生更广泛的影响，MoleculariumTM是一种结合了天文馆和虚拟太空之旅的产品，将观众带入原子和分子的世界。拟议研究的更广泛的社会成果包括开发下一代含糖药物，以及设计低成本、便携式芯片实验室系统，能够为护理点测试、研究和药物发现提供快速自动化生化分析。

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.