Nonlinear Imaging Methods for High-Throughput Protein and Drug Screening

用于高通量蛋白质和药物筛选的非线性成像方法

• 批准号: 8069282
• 负责人: JOHN C CONBOY
• 金额: $ 25.82万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8069282
• 关键词: ASIC channel; Accounting; Address; Adsorption; Affinity; Area; Behavior; Binding; Biological; Biological Assay; Biological Availability; Biological Models; Chemistry; Collaborations; DNA Microarray Chip; Detection; Development; Differential Scanning Calorimetry; Disadvantaged; Drug Compounding; Drug Delivery Systems; Drug Interactions; Electronics; Engineering; Environment; Equilibrium; Evaluation; Extravasation; Family; Fluorescence Microscopy; Frequencies; Funding; Gel; Generations; Goals; Grant; Health; Health Sciences; Heating; Image; Investigation; Ion Channel; Ions; Kinetics; Label; Light; Lipid Bilayers; Lipids; Liquid substance; Marines; Measures; Membrane; Membrane Lipids; Membrane Proteins; Methods; Microscope; Modeling; Molecular Weight; Optical Methods; Optics; Pathway interactions; Pharmaceutical Preparations; Phase; Phase Transition; Preclinical Drug Evaluation; Property; Proteins; Research; Research Design; Resolution; Sampling; Screening procedure; Signal Transduction; Solutions; Specificity; Spectrum Analysis; Structure; Sum; Surface; Techniques; Technology; Testing; Thermodynamics; Toxin; Universities; Utah; Vesicle; analog; analytical method; analytical tool; base; drug candidate; high throughput screening; imaging modality; inhibitor/antagonist; medical schools; membrane model; novel; patch clamp; receptor; reconstitution; response; second harmonic; small molecule; spectroscopic imaging; success; tool; ultraviolet

DESCRIPTION (provided by applicant): There is a growing need for analytical methods to identify and quantify the interaction of low molecular weight drug compounds with biological membranes. The use of lipid bilayer arrays has the potential to provide a cheap and efficient high- throughput analytical method capable of addressing these issues. There are several key issues which need to be resolved if lipid microarrays are to achieve the level of success obtained with DNA microarrays assays. Chief among these is the need for a noninvasive method to detect drug association to the lipid microarray surface and the subsequent perturbation of the lipid matrix due to drug interaction. The nonlinear techniques of ultraviolet-visible sum-frequency generation (UV-Vis SFG) and infrared- visible sum-frequency generation (IR-Vis SFG) imaging may hold the answer to this problem. Several attributes of nonlinear imaging make it a promising method for detecting drug-membrane interaction on microarrays, including the ability to quantify the detected signal, the high optical resolution and the inherent surface specificity. The crucial first steps to implementing UV-Vis SFG and IR-Vis SFG imaging for the investigation of drug association on lipid microarrays have already been achieved. Specific Aim #1 will focus on the development of UV-Vis SFG for the detection of low molecular weight molecules in planar supported lipid bilayers (PSLBs). The use of UV- Vis SFG for measuring protein adsorption to membranes, which is an extension of the goals of the previous grant period, will also be pursued. The nonlinear spectral response from several model protein and drug targets will be investigated in an attempt to more fully understand the factors controlling their detection by UV-Vis SFG in Aim #1. The application of UV-Vis SFG for the screening of potential ion channel inhibitors is explored in Aim #2. Examination of the influence of drugs on the thermotropic phase transition and phase behavior of lipid membranes by IR-Vis SFG spectroscopy and IR- Vis SFG high-throughput imaging is explored in Aim #3. These studies are designed to demonstrate the utility of nonlinear imaging methods in conjunction with micropatterned fluid lipid bilayer arrays for high-throughput drug screening applications. PUBLIC HEALTH RELEVANCE: Membrane proteins, either associated or integral, account for up to two thirds of known drug targets. The partitioning of drugs into the hydrophobic environment of the lipid membrane controls the bioavailability of these materials. The goals of the proposed studies are to develop analytical tools for the noninvasive, specific spectroscopic detection of proteins and small molecule (drug) interactions with biological membranes. The practical implementation of these technologies will have a significant impact on the pharmacological screening of drug candidates, and the investigation of drug-membrane interactions.

描述（由申请人提供）：越来越需要分析方法来识别和量化低分子量药物化合物与生物膜的相互作用。脂质双层阵列的使用有可能提供一种能够解决这些问题的廉价且高效的高通量分析方法。如果脂质微阵列要达到 DNA 微阵列检测的成功水平，则需要解决几个关键问题。其中最主要的是需要一种非侵入性方法来检测药物与脂质微阵列表面的关联以及随后由于药物相互作用而引起的脂质基质的扰动。紫外可见和频生成（UV-Vis SFG）和红外可见和频生成（IR-Vis SFG）成像的非线性技术可能会解决这个问题。非线性成像的几个属性使其成为检测微阵列上药物-膜相互作用的有前途的方法，包括量化检测到的信号的能力、高光学分辨率和固有的表面特异性。实施 UV-Vis SFG 和 IR-Vis SFG 成像以研究脂质微阵列药物关联的关键第一步已经实现。具体目标#1 将重点开发 UV-Vis SFG，用于检测平面支持的脂质双层 (PSLB) 中的低分子量分子。还将继续使用 UV-Vis SFG 来测量蛋白质对膜的吸附，这是上一个资助期目标的延伸。我们将研究多个模型蛋白质和药物靶标的非线性光谱响应，以尝试更全面地了解目标 #1 中控制 UV-Vis SFG 检测的因素。目标 #2 探讨了 UV-Vis SFG 在筛选潜在离子通道抑制剂中的应用。目标 #3 探讨了通过 IR-Vis SFG 光谱和 IR-Vis SFG 高通量成像检查药物对脂质膜热致相变和相行为的影响。这些研究旨在证明非线性成像方法与微图案流体脂质双层阵列相结合在高通量药物筛选应用中的实用性。公共卫生相关性：膜蛋白，无论是相关蛋白还是整体蛋白，占已知药物靶点的三分之二。药物在脂质膜的疏水环境中的分配控制着这些材料的生物利用度。拟议研究的目标是开发用于蛋白质和小分子（药物）与生物膜相互作用的非侵入性、特异性光谱检测的分析工具。这些技术的实际应用将对候选药物的药理学筛选以及药物与膜相互作用的研究产生重大影响。