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.
| Nagappayya, Sahana K; Pohl, Nicola L B (2012) Production of fluorous-based microarrays with uncharged carbohydrates. Methods Mol Biol 808:149-53 |
| Collet, Beatrice Y M; Nagashima, Tadamichi; Yu, Marvin S et al. (2009) Fluorous-based Peptide Microarrays for Protease Screening. J Fluor Chem 130:1042-1048 |
| Park, Gisun; Ko, Kwang-Seuk; Zakharova, Aleksandra et al. (2008) Mono- Vs. Di-flourous tagged Glucosamines for Iterative Oligosaccharide Synthesis. J Fluor Chem 129:978-982 |
