Fluorous Solution-Phase Synthesis of Peptides and Oligosaccharides

肽和寡糖的氟溶液相合成

• 批准号: 7908263
• 负责人: NICOLA L. B. POHL
• 金额: $ 49.56万
• 依托单位: FLUOROUS TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-04 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7908263
• 关键词: Acquired Immunodeficiency Syndrome; Anthrax disease; Antibodies; Attention; Automation; Binding; Biological; Biological Assay; Biological Process; Biopolymers; Carbohydrates; Carrier Proteins; Chromatography; Communities; Concanavalin A; 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; 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; 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.

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.基于在I阶段期间提出的实验的成功，在II期时期提出了进一步开发基于荧光的肽和寡糖合成和微阵列形成的实验。赠款的具体目的是：1）开发基于荧光的肽微阵列。将准备一个新的荧光连接器（2-氯二甲易甲甲），并将开发在N-或C末端制备带有荧光标签的肽的一般程序。将展示基于荧光的肽微阵列在酶和蛋白质肽结合测定中的效用。 2）基于荧光的碳水化合物合成自动化。使用市售的溶液相合成器，将开发自动荧光的碳水化合物合成方案。 3）开发自动合成的碳水化合物的纯化条件。将开发净化和分析由自动协议合成的碳水化合物的一般程序。 4）演示混合肽/碳水化合物微阵列。将在荧光素玻璃板上印刷荧光素的肽和单糖，并与荧光素异硫氰酸酯标记的Jack Bean凝集素浓蛋白A（FITC-CON A）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来说是有价值的，尤其是对于DUG，诊断和疫苗开发而言。本应用程序中提出的工作是为提供目前无法理解和改善人类健康状况的研究社区工具迈出的重要一步。通过使用单个化学惰性标签，将制造，纯化并固定重要的生物分子，例如肽和碳水化合物，以进行进一步研究。