Fluorous Solution-Phase Synthesis of Peptides and Oligosaccharides

肽和寡糖的氟溶液相合成

• 批准号: 7489863
• 负责人: NICOLA L. B. POHL
• 金额: $ 43.28万
• 依托单位: FLUOROUS TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7489863
• 关键词: Acquired Immunodeficiency Syndrome; Anthrax disease; Antibodies; Attention; Automation; Binding; Biological; Biological Assay; Biological Process; Biopolymers; Carbohydrates; Carrier Proteins; Chromatography; Class; Communities; Concanavalin A; Condition; Coupling; Development; Diagnostic; Enzymes; Exhibits; Fluorescein; Fluoresceins; Glass; Glycobiology; Glycoconjugates; Glycolipids; Glycopeptides; Glycoproteins; Goals; Grant; Health; Human; Immune; Inflammation; Influenza; Iowa; Isothiocyanates; Label; Lectin; Legal patent; Malaria; Malignant Neoplasms; Methods; Monosaccharides; Neoplasm Metastasis; Oligosaccharides; Pattern; Peptide Synthesis; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Play; Printing; Procedures; Proteins; Protocols documentation; Range; Reaction; Reporting; Research; Rest; Role; S Phase; Scientist; Screening procedure; Slide; Solid; Solutions; Source; Surface; System; Techniques; Technology; Testing; Time; Universities; Vaccines; Work; analog; analytical tool; base; bean; cancer cell; desire; fluorescein isothiocyanate-concanavalin A; immunogenicity; improved; pathogen; research study; success; tool; vaccine development

DESCRIPTION (provided by applicant): The long-term goal of the project is to develop new methods for (1) preparative scale syntheses of high purity peptides and oligosaccharides utilizing some of Fluorous Technologies' (FTI) proprietary techniques - fluorous tagging and automated fluorous solid phase extraction, and for (2) direct arraying of fluorous-tagged compounds based on technology developed and patented by Iowa State University. Based on the success of experiments proposed during the Phase I period, further development of fluorous-based peptide and oligosaccharide synthesis and microarray formation are proposed for the Phase II period. The specific aims of the grant are: 1) Development of fluorous-based peptide microarrays. A new fluorous linker (2-chlorotrityl) will be prepared, and general procedures to prepare peptides with fluorous tag at either N- or C-terminus will be developed. The utility of fluorous-based peptide microarrays in enzyme and protein-peptide binding assays will be demonstrated. 2) Automation of fluorous-based carbohydrate synthesis. Using a commercially available solution-phase synthesizer, automated fluorous-based carbohydrate synthesis protocols will be developed. 3) Development of purification conditions for carbohydrates derived from automated synthesis. General procedures to purify and analyze carbohydrates synthesized by automated protocol will be developed. 4) Demonstration of a mixed peptide/carbohydrate microarray. Fluorous-tagged peptides and monosaccharides will be printed on a fluorous glass plate, and binding to the fluorescein isothiocyanate- labeled jack bean lectin concanavalin A (FITC-con A) will be studied. We expect the developed methods and commercial tools to be valuable to biologists, chemists, and pharmaceutical scientists, especially for dug, diagnostics, and vaccine development. The work proposed in this application represents a significant step toward providing the research community tools not currently available to understand and improve the human health condition. Through the use of a single chemically inert tag, important biomolecules such as peptides and carbohydrates will be made, purified and immobilized for further study.

描述（由申请人提供）：该项目的长期目标是开发新的方法：(1)利用氟技术公司（FTI）的一些专利技术——氟标记和自动氟固相萃取——制备高纯度多肽和低聚糖的大规模合成，以及(2)基于爱荷华州立大学开发和专利的技术直接排列氟标记化合物。基于第一阶段实验的成功，建议在第二阶段进一步发展含氟多肽和低聚糖的合成以及微阵列的形成。该基金的具体目标是：1)开发基于氟的肽微阵列。本文将制备一种新的含氟连接剂（2-氯三硝基），并将研究制备N端或c端含氟肽的一般方法。氟基肽微阵列在酶和蛋白肽结合分析中的应用将被证明。2)含氟碳水化合物合成自动化。使用市售的液相合成器，将开发基于氟的碳水化合物自动合成方案。3)自动合成碳水化合物纯化条件的研究。将开发一般程序来纯化和分析由自动化协议合成的碳水化合物。4)多肽/碳水化合物混合芯片的演示。将含氟标记的多肽和单糖印刷在含氟玻璃板上，并研究与异硫氰酸荧光素标记的豆角凝集素-豆蛋白a （FITC-con a）的结合。我们期望开发的方法和商业工具对生物学家、化学家和制药科学家有价值，特别是在挖掘、诊断和疫苗开发方面。本申请中提出的工作代表了向提供研究界目前无法获得的工具来了解和改善人类健康状况迈出的重要一步。通过使用单一的化学惰性标签，重要的生物分子如肽和碳水化合物将被制造、纯化和固定化以供进一步研究。