Development of Atomic Force Microscopic Instrumentation for the Detection of Chemical and Biological Forces

该提案旨在建立一个跨学科的研究计划，在爱荷华州州立大学（ISU）在生物分子力的检测使用原子力显微镜（AFM）的原则。这一目标将通过在ISU成立一个合作研究小组来实现，该小组由动物学遗传学系和化学系的科学家组成，逐步发展到包括与ISU其他部门的研究人员合作。研究的中心焦点将是检测和测量在相对简单的（例如，氨基酸）和更复杂的（例如，短肽、蛋白质/核酸、抗体-抗原）生物分子系统。从这些调查中获得的信息将提供重要的洞察力的分子识别，自组装和折叠的生物分子系统的结合模式。 此外，这些研究将加强新兴分子力检测技术在生物分析诊断中的应用。 该计划的开发分为三个相互关联的项目，所有这些项目都试图利用AFM及其变体在纳米长度尺度上探测界面结构的能力。这三个项目是： 项目1：头端-基底改性方法。该项目的重点是开发新的，或现有的，用于AFM针尖的化学和生物改性的方法的扩展。 还将设计用于构建空间图案化表面的方法，作为用于制备不同分子种类的小的局部阵列的平台。 项目2：分子力相互作用。本项目使用项目1中创建的材料，通过AFM力表征研究生物材料的分子水平相互作用。该项目从各种接触液体（例如，有机溶剂和生物介质）的模型化学系统（例如，- CO2 H，-CH 3），发展到越来越复杂的系统（例如，氨基酸，短肽），并以研究超分子结构域之间的生物分子力相互作用（例如，亮氨酸拉链）和整个分子（例如，抗体/抗原复合物）。 项目3：生物分子力显微镜（BMFM）。该项目需要建造第一代AFM，专门用于检测和测量生物材料之间的力。BMFM将基于分子力检测的既定设计原则和项目1和2的研究结果。 为执行上述项目，需要提供资金，用于购置和建造仪器，并为与仪器的设计、开发、操作和维护有关的人员和用品提供部分支助。 作为机构费用分摊的一部分，执行支助股将为仪器设备以及为制定这一方案而开展的若干研究和培训活动提供支助。这种支持远远超过了NSF建议的50%的成本份额，表明ISU对该计划的启动和发展的高水平承诺。

This proposal seeks to establish an interdisciplinary research program at Iowa State University (ISU) in the detection of biomolecular forces using the principles of atomic force microscopy (AFM). This goal will be accomplished by the formation of a collaborative research group at ISU that is composed of scientists from the Department of Zoology & Genetics and the Department of Chemistry, evolving to include collaborations with investigators in other ISU departments. The central focus of research will be the detection and measurement of the binding forces operative in the interactions between relatively simple (e.g., amino acids) and more complex (e.g., short peptides, protein/nucleic acid, antibody-antigen) biomolecular systems. The information gained from these investigations will provide important insight into the binding modes related to the molecular recognition, self-assembly, and folding of biomolecular systems. Furthermore, these studies will enhance the implementation of emerging molecular force detection techniques in bioanalytical diagnostics. The development of this program is divided into three interrelated projects, all of which seek to take advantage of the ability of AFM and its variants to probe interfacial architectures at nanometer length scales. The three projects are: Project 1: Tip-Substrate Modification Methods. This project focuses on the development of new, or the extension of existing, methods for the chemical and biological modification of AFM tips. Approaches will also be devised for the construction of spatially patterned surfaces as platforms for the preparation of small, localized arrays of different molecular species. Project 2: Molecular Force Interactions. This project investigates the molecular level interactions of biological materials through AFM-force characterizations using the materials created in Project 1. The project starts with characterizations under a variety of contacting liquids (e.g., organic sol vents and biological media) of model chemical systems (e.g., -CO2H, -CH3), progresses to increasingly complex systems (e.g., amino acids, short peptides), and culminates with studies of the biomolecular force interactions between supermolecular domains (e.g., the leucine zipper) and whole molecules (e.g., antibody/antigen complexes). Project 3: The BioMolecular Force Microscope (BMFM). This project entails the construction of a first generation AFM that is designed specifically for the detection and measurement of forces between biological materials. The BMFM will be based on established design principles for molecular force detection and on the findings in Projects 1 and 2. To carry out the above projects, funding is requested for instrumentation acquisition and construction, and for partial support of personnel and supplies associated with the design, development, operation, and maintenance of the instrumentation. As part of the institutional costsharing, ISU will provide support for instrumentation and for several of the research and training activities for the development of this program. This support extends well past the recommended 50% cost share by NSF, demonstrating the high level of commitment by ISU to the initiation and growth of the program.