Fluorous Solution-Phase Synthesis of Peptides and Oligosaccharides

描述(由申请人提供)：该项目的长期目标是开发新的方法，用于(1)利用一些氟技术(FTI)的专利技术--氟标记和自动氟固相萃取--制备规模的高纯度多肽和寡糖，以及(2)基于爱荷华州立大学开发和专利的技术直接对带有氟标记的化合物进行排列。在第一阶段实验取得成功的基础上，建议第二阶段进一步开展含氟多肽和低聚糖的合成和微阵列的形成。该赠款的具体目标是：1)开发基于氟的多肽微阵列。将制备一种新的含氟连接物(2-氯三苯基)，并将制定在N-端或C-端带有氟标记的多肽的一般制备程序。将展示基于氟的多肽微阵列在酶和蛋白质-多肽结合分析中的应用。2)含氟碳水化合物合成的自动化。利用商业上可用的溶液相合成器，将开发基于氟的碳水化合物自动合成方案。3)开发来自自动合成的碳水化合物的纯化条件。将开发提纯和分析由自动化程序合成的碳水化合物的通用程序。4)展示了一种多肽/碳水化合物混合芯片。氟标记的多肽和单糖将被印刷在氟玻璃板上，并将研究与荧光素异硫氰酸酯标记的菜豆凝集素刀豆凝集素刀豆蛋白A(FITC-conA)的结合。我们希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。这项申请中提出的工作代表着朝着向研究社区提供目前尚不具备的工具来了解和改善人类健康状况迈出了重要的一步。通过使用单一的化学惰性标签，将制造、纯化和固定化重要的生物分子，如多肽和碳水化合物，以供进一步研究。

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.