Biacore 3000 System

描述(由申请人提供)：需要一种商业上可用的仪器，用于大分子络合物的定量表面等离子体共振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，这些研究共享基本的长期目标，即了解构成生物和医学相关相互作用基础的化学和物理原理。这里正在研究的广泛的系统目前由于需要为每个系统开发专门的结合分析而受到阻碍。此外，目前的分析方法具有有限的动态范围和/或令人望而却步的材料要求，并且产生的信息比拟议的设备提供的信息更有限。该提案记录了用户访问所建议的仪器将如何克服目前使用的方法的以下限制：缺乏监测结合事件的非干扰实验观察物；大分子样品的数量和/或浓度高得令人望而却步；观察的灵敏度和动态范围受到严格限制；以及多样品分析的限速吞吐量。据报告，初步研究表明该文书对其预定目的的适用性以及有效执行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教授配体结合分析的基本原理和实践提供支持。

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.