Biacore 3000 System

描述（由申请人提供）：需要一种商用仪器，用于大分子复合物的表面等离子体共振定量检测和分子组分的回收。该仪器通过产生的质量变化直接检测结合事件。它在校园的存在将极大地促进正在进行的、由美国国立卫生研究院资助的基础研究，这些研究的基本长期目标是理解生物学和医学相关相互作用背后的化学和物理原理。目前，由于需要为每一种系统开发专用的结合试验，正在研究的广泛系统受到阻碍。此外，目前的测定法具有有限的动态范围和/或禁制性材料要求，并且产生的信息比拟议设备提供的信息更有限。该提案记录了用户如何访问所提议的仪器将克服当前采用的方法的以下限制：缺乏监测绑定事件的非干扰实验观察；大分子样品的数量和/或浓度过高；观测灵敏度和动态范围受到严重限制；以及多样品分析的限速吞吐量。报告的初步研究表明，该文书对其预期目的的适用性以及有效实施该文书所需的专门知识。该仪器还将为研究生和高级本科生提供配体结合分析的基本原理和实践指导。

DESCRIPTION (provided by applicant): A commercially available instrument for quantitative surface plasmon resonance detection of macromolecular complexes and recovery of molecular components is requested. This instrument detects binding events directly through the resulting change in mass. Its presence on campus will greatly facilitate ongoing, NIH-funded basic investigations that share the fundamental long-term goal of understanding the chemical and physical principles that underlie biologically and medically relevant interactions. A wide range of systems under study here are presently hampered by the need for developing a dedicated binding assay for each one. In addition, current assays have limited dynamic range and/or prohibitive materials requirements, and yield more limited information than is available from the proposed equipment. This proposal documents how user access to the proposed instrumentation will overcome the following limitations of presently employed methods: lack of non-perturbing experimental observables that monitor binding events; prohibitively high amounts and/or concentrations of macromolecular samples; severely restricted sensitivity and dynamic range of observations; and rate-limiting throughput for multi-sample analysis. Preliminary studies are reported that demonstrate the applicability of the instrument for its intended purposes and the expertise required for its effective implementation. The instrument will also support instruction in the fundamental principles and practice of ligand-binding analysis for graduate and advanced undergraduate students.