Digital Microfluidic Artificial Golgi for Glycan Synthesis

用于聚糖合成的数字微流体人工高尔基体Jonathan DordickRensselaer Polytechnic InstituteCBET-0730817糖基化是蛋白质翻译后最重要的修饰，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理解，并且使得必需治疗剂的受控合成困难且昂贵。因此，迫切需要对高尔基体内聚糖生物合成的基本理解，这将导致人工途径的发展，以合成，修饰，并最终大规模合成聚糖和糖蛋白。本研究的目标是通过数字微流控系统（DMFS）进行硫酸乙酰肝素/肝素的受控生物催化合成，DMFS是一类新型的芯片实验室系统，可在电极阵列上操纵离散的化学品液滴。智力优势：这项研究将导致在高尔基体的糖基化的基本理解，并通过检查精确控制的酶催化的空间和时间参数的变化来模拟这些过程。作为一个模型，硫酸乙酰肝素/肝素的体外合成将代表人工高尔基体的第一个演示，可能是在各种各样的治疗性糖蛋白的合成的实用程序。精确的控制算法将被开发，以实现有效的糖基化，并将建立一个全面的框架来设计流体的运动，保留和分离，就像在高尔基体细胞器。更广泛的影响：结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。将该项目与RPI正在进行的MoleculariumTM开发工作相结合将产生更广泛的影响，MoleculariumTM是一种结合了天文馆和虚拟太空旅行的产品，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聚糖的药物，以及设计低成本、便携式芯片实验室系统，该系统能够进行快速自动生化分析，用于即时检测、研究和药物发现。

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.