Static and dynamic light scattering instrumentation for macromolecular characteri

描述（由申请人提供）：本申请申请要求资助用于表征大分子尺寸、形状、低聚状态和溶液行为的最先进的光散射（LS）装置。仪器将由Wyatt Technology Corporation DAWN HELEOS MALS（多角度静态光散射）系统、WyattQELS（准弹性光散射，即，动态光散射）仪器，以及与Agilent HPLC连接的Optilab雷克斯折射率检测器，用于对通过尺寸排阻和其他形式的色谱法分离的样品进行在线、真实的时间LS分析。这些设备将提供康奈尔大学目前无法提供的分析能力，这对十几个NIH资助的赠款至关重要。拟议的仪器将补充康奈尔大学现有的仪器，特别是在分析完整和交换蛋白质复合物，蛋白质-蛋白质/配体相互作用和探测蛋白质构象方面。蛋白质大小和形状的准确评估是分子表征的关键第一步。此外，关联系统是许多信号转导和生物调节机制的基础。从复合物的形成中描述构象变化变得越来越重要。通过与MALS/DLS相结合的分离技术对异质样品进行仔细分析可以以时间和成本有效的方式实现这些目标。此外，许多其他形式的结构分析，如X射线晶体学和NMR光谱学，依赖于确定样品是单分散和均匀的条件。MALS/DLS分析将大大提高和加速我们筛选样品进行结晶试验和其他生物物理实验的能力。该仪器的特点是：1）可同时进行18个角度的静态光散射测量，直接测量分子量和绝对分子量; 2）在进行静态LS测量的波长下，可测定溶液的折射率（n）和折射率随浓度的变化（dn/dc）; 3）可同时测定强度波动的扩散系数; 4）先进的分析软件，提供每次测量的标准偏差; 5）批量或流动池分析，用于检测小样品或耦合的HPLC洗脱液。该仪器的获得将对十几个NIH资助的研究项目产生重要影响，并将改善康奈尔大学的研究基础设施以及学生和研究人员的教育/培训计划。它将加速和促进康奈尔大学三个学院与生命科学研究相关的广泛研究项目，并有助于保持康奈尔大学的竞争地位。

DESCRIPTION (provided by applicant): This application requests funds for state-of-the-art Light Scattering (LS) devices for the characterization of macromolecular size, shape, oligomeric state and solution behavior. The instrumentation will consist of a Wyatt Technology Corporation DAWN HELEOS MALS (Multi-angle static light scattering) system, a WyattQELS (Quasi-Elastic-Light-Scattering, i.e., dynamic light scattering) instrument, and a Optilab rEX refractive index detector interfaced with an Agilent HPLC for in-line, real time LS analysis of samples separated by size exclusion and other forms of chromatography. This equipment will provide analytical capabilities currently not available at Cornell, essential for over a dozen active NIH supported grants. The proposed instrument will complement existing instrumentation at Cornell, particularly with respect to analyzing intact and exchanging protein complexes, protein-protein/ligand interactions and probing protein conformation. Accurate assessment of protein size and shape is a critical first step in molecular characterization. Moreover, associating systems underlie many mechanisms of signal transduction and biological regulation. It is becoming increasingly important to delineate conformational change from complex formation. Careful analyses of heterogenous samples by separation techniques combined with MALS/DLS can achieve these goals in a time and cost effective manner. Furthermore, many other forms of structural analysis such as x-ray crystallography and NMR spectroscopy rely on identifying conditions where samples are monodisperse and homogeneous. MALS/DLS analysis will greatly enhance and accelerate our ability to screen samples for crystallization trials and other biophysical experiments. The particular strengths of the proposed instrument are 1) Direct and absolute molecular weight measurements from simultaneous 18 angle static light scattering measurements; 2) Evaluation of the solution refractive index (n) and the change in refractive index with concentration (dn/dc) at the wavelength where static LS measurements are made; 3) Simultaneous assessment of diffusion coefficients from intensity fluctuations; 4) Sophisticated analysis software that provides standard deviations on each measurement; 5) Batch or flow cell analysis for examination of small samples or coupled HPLC eluates. The acquisition of this instrument will have an important impact on more than a dozen NIH-funded research projects, and will improve the research infrastructure of Cornell University and the education/training programs of students and researchers. It will accelerate and facilitate a wide range of research projects of Cornell faculty across three colleges relevant to life sciences research, and help to maintain Cornell's competitive position.