Refinement and implementation of the automated oligosaccharide synthesizer

自动化寡糖合成仪的完善和实现

• 批准号: 9346826
• 负责人: ALEXEI V DEMCHENKO
• 金额: $ 7.51万
• 依托单位: UNIVERSITY OF MISSOURI-ST. LOUIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9346826
• 关键词: Academia; Area; Attention; Automation; Biochemistry; Biological; Biological Sciences; Carbohydrate Chemistry; Carbohydrates; Chemicals; Chemistry; Communities; Complex; Computer Interface; Computer software; Development; Diagnostic; Discipline; Disease; Equipment; Evolution; Genomics; Goals; Government; Health; High Pressure Liquid Chromatography; Human; Individual; Industry; Knowledge; Laboratories; Left; Libraries; Life; Liquid substance; Manuals; Medical; Medicine; Methods; Microarray Analysis; Mind; Monitor; Nature; Oligosaccharides; Organic Synthesis; Organism; Plant Resins; Polymers; Polysaccharides; Preparation; Procedures; Proteomics; Public Health; Reaction Time; Reagent; Reporting; Research; Research Personnel; Research Project Grants; Role; S Phase; Science; Scientist; Sialic Acids; Societies; Solid; Solvents; Specialist; Structure; System; Technology; Therapeutic; Therapeutic Agents; Thioglycosides; Time; Training; base; computer program; cost; detector; dietary supplements; experience; glycosylation; innovation; interest; programs; research and development; symposium; therapeutic vaccine; time use; user-friendly

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.

寡糖自动合成仪的改进与实现 项目摘要 碳水化合物形成了所有生物体的基础，因此，在自然界中无处不在， 生物活性化合物和在医学中作为治疗剂。尽管人们一直对 碳水化合物的合成，以及这些分子固有的复杂性， 许多科学家的注意，用于分离、化学或酶促合成、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要求开发快速，高效， 操作简单。虽然这是一项艰巨的任务，但这项建议将侧重于 引入一个负担得起的和可访问的自动化平台，使专家和非专家都能 专家进行聚糖的合成。 目前用于寡糖合成的自动化方法是高度复杂的，操作上复杂。 复杂，需要大量的用户知识。相比之下，高效液相色谱（HPLC） Demchenko和Stine介绍的基于设备的自动化代表了一种高度可访问的方法， 合成.这是因为许多糖科学家都有HPLC的操作知识，并且很容易获得HPLC 设备.自动化合成不仅通过使用化学试剂递送所有试剂而提供操作简单性， 标准自动进样器，而且还可以使用检测器方便地实时监测每个步骤的反应， 标准计算机软件和接口。我们使用更少的反应物、试剂和 溶剂比手工固相合成。在有希望的初步结果的基础上，该建议旨在 生成一个用户友好的通用平台，用于自动化和转化聚糖合成。 虽然所提出的自动化方法有可能彻底改变聚糖合成，但实际上， 固相合成方面需要改进。这项建议分为四个具体目标。第一 三个目标致力于改进固相合成的固有局限性：1）新的负担得起的 固相合成的树脂和通用连接剂; 2）催化和立体控制的新概念 糖基化;和3）用于简化获得结构单元的新方法。第四个目标是致力于 实施自动进样器作为寡糖合成自动化的模式。完成后 在拟议的研究中，我们期望获得一个可靠和简单的平台， 寡糖合成许多种类的化合物可以使用固体载体获得;因此， 预计拟议的研究将对许多科学家具有宝贵的方法和实际用途。调查人员 能够使用标准HPLC设备的人应该能够进行聚糖库的自动化合成 使用我们的试剂盒或与现有的方法和设备轻松接口。合成 碳水化合物和其他类别的生物分子使用这个用户友好的自动化平台将加速 发现在许多科学学科，并可以显着影响技术，社会，经济， 公共卫生