Refinement and implementation of the automated oligosaccharide synthesizer

全自动低聚糖合成仪的改进与实现 项目摘要 碳水化合物构成了所有生命有机体的基础，因此在自然界中普遍存在于 具有生物活性的化合物，在医学上用作治疗药物。尽管一直以来人们对 碳水化合物的合成，以及这些分子固有的复杂性始终捕捉到 许多科学家的关注，分离的方法，化学或酶合成，分析和 碳水化合物的应用仍然很麻烦。不断增加的时间限制和成本限制 糖科学和糖技术的研究和发展呼唤快速、高效和 操作简单的程序。虽然这是一项艰巨的任务，但这项提案将侧重于 推出经济实惠且易于访问的自动化平台，使专家和非 专家进行多糖的合成。 目前用于低聚糖合成的自动化方法在操作上是高度复杂的 复杂，并且需要大量的用户知识。相比之下，高效液相色谱(HPL C) Demchenko和Stine引入的基于设备的自动化代表了一种高度可访问的方法 综合。这是因为许多血糖科学家具有高效液相色谱的操作知识，并且很容易获得。 设备。自动化合成不仅提供了操作上的简单性，通过使用 标准自动进样器，还可以使用检测器和 标准的计算机软件和界面。我们使用更少的反应物、试剂和 溶剂比人工固相合成要好。在有希望的初步结果的基础上，这项提议旨在 创建一个用户友好的通用平台，用于自动化和变革性的多糖合成。 虽然拟议的自动化方法有可能彻底改变多糖的合成，但几乎每一个 固相合成方面需要改进。这项提议被组织成四个具体目标。第一 三个目标致力于改善固相合成的固有局限性：1)新的负担得起的 树脂和固相合成的通用连接物；2)催化和立体控制的新概念 糖基化；以及3)简化获取积木的新方法。第四个目标是致力于 实现自动进样器作为低聚糖合成的自动化模式。完工后 在建议的研究中，我们预计已经获得了一个可靠和简单的自动化平台 低聚糖合成。使用固体载体可以获得许多种类的化合物；因此， 所提出的研究有望对许多科学家具有有价值的方法论和实用价值。调查人员 有了标准的高效液相设备，应该能够进行糖链文库的自动合成 使用我们的试剂盒，或与其现有方法和设备轻松对接。一种新的合成方法 碳水化合物和其他种类的生物分子使用这个用户友好的自动化平台将加速 在许多科学学科中的发现，并可显著影响技术、社会、经济和 公共卫生。

Refinement and Implementation of the Automated Oligosaccharide Synthesizer Project Summary Carbohydrates form the basis of all living organisms and, as a consequence, are ubiquitous both in nature as biologically active compounds and in medicine as therapeutics. Although there has been continued interest in the synthesis of carbohydrates, and the inherent complexity of these molecules consistently captures the attention of many scientists, methods for the isolation, chemical or enzymatic synthesis, analysis, and application of carbohydrates remain cumbersome. Ever-increasing time constraints and cost limitations on research and development in glycoscience and glycotechnology call for the development of rapid, efficient and operationally simple procedures. While this represents an immense task, this proposal will focus on the introduction of an affordable and accessible automation platform that will enable both specialists and non- specialists to perform the synthesis of glycans. Current automated methods for the synthesis of oligosaccharides are highly sophisticated, operationally complex, and require significant user know-how. By contrast, high performance liquid chromatography (HPLC) equipment-based automation introduced by Demchenko and Stine represents a highly accessible method of synthesis. This is because many glycoscientists have operational knowledge of, and easy access to, HPLC equipment. The automated synthesis offers not only operational simplicity by delivering all reagents using standard autosamplers, but also convenient real-time reaction monitoring of every step using detectors and standard computer software and interface. We achieved faster reaction times using less reactant, reagent, and solvent than manual solid-phase synthesis. Building upon promising preliminary results, this proposal aims to generate a user-friendly, universal platform for automated and transformative synthesis of glycans. While the proposed automation approach has the potential to revolutionize glycan synthesis, practically every aspect of solid phase synthesis needs to be refined. This proposal is organized into four specific aims. The first three aims are dedicated to the refinement of inherent limitations of solid phase synthesis: 1) new affordable resins and the universal linker for solid phase synthesis; 2) new concepts for catalytic and stereocontrolled glycosylation; and 3) new methods for streamlined access to building blocks. The fourth aim is dedicated to the implementation of autosamplers as a mode for automation of the oligosaccharide synthesis. Upon completion of the proposed studies, we expect to have acquired a reliable and simple platform for automated oligosaccharide synthesis. Many classes of compounds can be obtained using solid supports; therefore, the proposed study is expected to be of valuable methodological and practical use to many scientists. Investigators with access to standard HPLC equipment should be able to perform automated synthesis of glycan libraries using our reagent kits or easily interface with their existing methods and equipment. Synthesis of carbohydrates and other classes of biomolecules using this user-friendly, automated platform will accelerate discovery in many scientific disciplines and can significantly impact technology, society, the economy, and public health.