DEVELOPMENT OF FORCE SPECTROSCOPY FOR GENERAL USE

描述(改编自申请人的摘要)：本报告的目标 建议是开发必要的工具和方法，以使 分子间和分子内力的直接机械测量 用户友好、价格低廉、适用范围广的分子分析工具 和高通量的分子筛选。这种新的方法基于 可测量原子力显微镜(AFM)的原理 皮牛顿范围内的力，并将这些力转换为宏观 信号。在建议的方案中，一种简单的超敏感和成本效益 将建造和测试F-SPEC装置。此设备将包括一个流 小室和用于暂时固定微粒的机制 允许快速、顺序地分析与 微粒。该系统将使用两个分子系统进行评估 以前由F-SPEC研究；亲和素/生物素和抗体/抗原分子 成对的。将进一步发展使用DNA-蛋白质和分子内的方法 DNA发夹分子系统。这些努力将导致产生 一种测量分子内部和分子间作用力的技术，它将具有 在许多生物医学重要领域的应用，包括基础 生物分子分析、信号转导、免疫学和免疫诊断学 表面的药理、药物发现和高通量筛选 固定化组合分子文库。

DESCRIPTION (Adapted from the applicant's abstract): The goal of this proposal is to develop the instrumentation and methodology necessary to make direct mechanical measurement of inter- and intra-molecular forces, F-SPEC, a user friendly, inexpensive and broadly applicable tool for molecular analysis and high throughput molecular screening. This novel approach is based on the principles of the atomic force microscope (AFM) which is capable of measuring forces in the pico Newton range and transducing these forces into macroscopic signals. In the proposed program, a simple ultra-sensitive and cost effective F-SPEC device will be constructed and tested. This device will include a flow chamber and a mechanism for temporary immobilization of microparticles to allow rapid, sequential analysis of molecular species tethered to the microparticles. The system will be evaluated using two molecular systems previously studied by F-SPEC; avidin/biotin and antibody/antigen molecular pairs. Methods will be further developed using DNA-protein and intramolecular DNA hairpin molecular systems. These efforts will result in the production of a technology for measuring forces in and between molecules, that will have applications in a large number of biomedically important areas including basic biomolecular analysis, signal transduction, immunology and immunodiagnostics, pharmacology, drug discovery and high throughput screening of surface immobilized combinatorial molecular libraries.