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.

人乳低聚糖化学合成的自动化 项目摘要 碳水化合物是所有生物体的基础，在自然界和医学中无处不在。 然而，用于化学或酶促合成碳水化合物的方法仍然繁琐。快速 糖科学研发的增长要求开发快速、有效和简单的聚糖分析方法， 合成.这项建议旨在满足这一需求，重点是推出一个负担得起的， 一个可访问的自动化平台，使专家和非专家都能进行综合， 聚糖 目前用于合成聚糖的方法是高度复杂的、操作复杂的，并且需要 重要的用户知识。相比之下，高效液相色谱设备以自动化为主 由Demchenko和Stine引入的高效液相色谱法（HPLC-A）代表了一种高度可及的合成方法， 许多科学家已经具有HPLC设备的操作知识并且容易获得HPLC设备。自动化 合成不仅通过使用标准HPLC组分递送所有试剂而提供操作简单性， 也便于对每一步反应进行实时监控。该提案旨在改进我们目前的HPLC-A 通过引入一个通用平台来实现聚糖的完全自动化合成。 我们的前两个目标致力于解决我们以前平台的关键弱点，我们的第三个目标将 将这些改进应用于人乳低聚糖（HMO）的合成。我们将致力于 用于聚糖固相合成的具有增强性能的整体载体（Aim A）。我们还将 开发专用的糖基化反应，并为我们的系统配备新的模块， 再循环效率，提高糖基化的再现性，减少对过量结构单元的需要， 并导致合成的所有步骤完全自动化（目标B）。然后我们将展示如何以及这些 战略调整可以应用于HMO的完全自动化的“按下按钮”生产， 其作为益生元、抗微生物剂和营养素在婴儿发育中具有关键作用（Aim C）。 在完成建议的研究后，我们预期已建立一个可靠和简单的平台， 全自动合成。能够使用标准HPLC设备的研究者应能够 使用我们的方法进行聚糖的自动合成。机器辅助合成确保了严格的 实验设计，以获得稳健的结果，消除变异性，并准确地重现实验 不同的用户多次。使用这种用户友好的自动化平台合成聚糖将 加速许多科学学科的发现，并对技术、社会、 经济和公共卫生。