Biacore 3000 System

描述（由申请人提供）：需要一种用于大分子复合物定量表面等离子体共振检测和分子组分回收的市售仪器。该仪器通过产生的质量变化直接检测结合事件。它在校园里的存在将极大地促进正在进行的，NIH资助的基础研究，这些研究共享的基本长期目标是理解生物和医学相关相互作用的化学和物理原理。目前，由于需要为每一种系统开发专用的结合测定法，因此在此研究的各种系统受到阻碍。此外，目前的测定具有有限的动态范围和/或禁止的材料要求，并且产生比从所提出的设备可获得的信息更有限的信息。该提案记录了用户如何访问拟议的仪器将克服目前采用的方法的以下限制：缺乏非扰动的实验观测值，监测结合事件;高得令人望而却步的量和/或浓度的大分子样品;严重限制的灵敏度和动态范围的观察;和速率限制多样品分析的吞吐量。报告的初步研究表明，该文书适用于其预期目的，并具备有效执行文书所需的专门知识。该仪器还将支持研究生和高级本科生配体结合分析的基本原理和实践的教学。

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.