Automation of the chemical synthesis of human milk oligosaccharides

人乳低聚糖化学合成的自动化

• 批准号: 10318919
• 负责人: ALEXEI V DEMCHENKO
• 金额: $ 30.82万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318919
• 关键词: Address; Area; Automation; Biological; Carbohydrate Chemistry; Carbohydrates; Chemicals; Collection; Communities; Complex; Computer software; Development; Discipline; Disease; Ensure; Equipment; Evolution; Experimental Designs; Goals; Health; High Pressure Liquid Chromatography; Human; Human Milk; Infant Development; Infant formula; Intervention; Knowledge; Lead; Length; Manuals; Medical; Medicine; Methods; Monitor; National Institute of General Medical Sciences; Nature; Nutrient; Oligonucleotides; Oligosaccharides; Organism; Peptides; Plant Resins; Polymers; Polysaccharides; Polystyrenes; Positioning Attribute; Preparation; Procedures; Production; Property; Protocols documentation; Public Health; Reaction; Reaction Time; Reagent; Reproducibility; Research; Research Personnel; Research Project Grants; Role; S phase; Science; Scientist; Societies; Solid; Specialist; Strategic Planning; System; Technology; Therapeutic; Time; Vaccines; Vision; Work; analytical method; analytical tool; antimicrobial drug; base; biological systems; chemical resource; chemical synthesis; computer program; design; dietary supplements; experimental study; glycosylation; improved; laboratory equipment; nanomaterials; prebiotics; programs; rapid growth; regenerative; research and development; user-friendly

Automation of the chemical synthesis of human milk oligosaccharides Project Summary Carbohydrates form the basis of all living organisms and are ubiquitous both in nature and in medicine. However, methods for the chemical or enzymatic synthesis of carbohydrates remain cumbersome. The rapid growth of R&D in glycoscience demands the development of rapid, efficient and simple procedures for glycan synthesis. This proposal seeks to meet this demand by focusing 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 methods for the synthesis of glycans are highly sophisticated, operationally complex, and require significant user know-how. By contrast, high performance liquid chromatography equipment-based automation (HPLC-A) introduced by Demchenko and Stine represents a highly accessible method of synthesis because many scientists already have operational knowledge of, and easy access to, HPLC equipment. Automated synthesis offers not only operational simplicity by delivering all reagents using standard HPLC components, but also convenient real-time reaction monitoring of every step. This proposal aims to improve our current HPLC-A setup by introducing a universal platform for completely automated synthesis of glycans. Our first two aims are dedicated to addressing key weaknesses in our previous platform, and our third aim will apply these improvements to the synthesis of human-milk oligosaccharides (HMO). We will develop dedicated monolithic supports with enhanced properties for solid phase synthesis of glycans (Aim A). We will also develop a dedicated glycosylation reaction and accessorize our system with new modules that will enhance the efficiency of recirculation, improve reproducibility of glycosylations, reduce the need for excess building block, and lead to complete automation of all steps of the synthesis (Aim B). We will then demonstrate how well these strategic adjustments can be applied to the completely automated, “press of a button” production of HMO, which have crucial roles as prebiotics, antimicrobial agents, and nutrients in infant development (Aim C). Upon completion of the proposed studies, we expect to have achieved a reliable and simple platform for completely automated synthesis. Investigators with access to standard HPLC equipment should be able to perform automated synthesis of glycans using our methods. Machine-assisted synthesis ensures rigorous experimental design to obtain robust results, to eliminate variability, and to accurately reproduce experiments multiple times by different users. Synthesis of glycans using this user-friendly, automated platform will accelerate discovery in many scientific disciplines and can significantly impact technology, society, the economy, and public health.

人奶寡糖化学合成的自动化 项目摘要 碳水化合物构成了所有生物体的基础，并且在自然界和医学上都是无处不在的。 然而，碳水合物化学或酶合成的方法仍然繁琐。快速 聚糖中研发的增长需要开发快速，高效和简单的过程 合成。该建议旨在通过关注负担得起的和 可访问的自动化平台将使专家和非专家们能够执行合成 聚糖。 当前合成聚糖的方法是高度复杂的，操作上复杂的，并且需要 重要的用户专有技术。相比之下，高性能液相色谱基于设备的自动化 （HPLC-A）由Demchenko和Stine引入 许多科学家已经对HPLC设备具有运营知识，并且可以轻松使用。自动化 综合不仅通过使用标准HPLC组件传递所有试剂来提供操作简单性，还提供 也方便每个步骤的实时反应监视。该建议旨在改善我们当前的HPLC-A 通过引入一个通用平台，用于完全自动化的聚糖平台。 我们的前两个目标致力于解决以前平台中的关键弱点，我们的第三个目标将 将这些改进应用于人乳寡糖（HMO）的合成。我们将发展敬业 具有增强性能的单层支持，用于聚糖的固相合成（AIM A）。我们也会 开发专用的糖基化反应，并使用新模块为我们的系统配附加，以增强 再循环效率，提高糖基化的可重复性，减少对超过构建块的需求， 并导致合成所有步骤的完整自动化（AIM B）。然后，我们将证明这些 战略调整可以应用于HMO的完全自动化的“按钮”制作， 在婴儿发育中具有至关重要的作用，作为益生元，抗菌剂和营养素（AIM C）。 拟议的研究完成后，我们希望获得一个可靠而简单的平台 完全自动合成。可以使用标准HPLC设备的调查人员应该能够 使用我们的方法对聚糖进行自动合成。机器辅助合成可确保严格 实验设计以获得强大的结果，以消除可变性并准确再现实验 不同用户多次。使用此用户友好，自动化平台合成聚糖 在许多科学学科中加速发现，并可能对技术，社会，社会， 经济和公共卫生。