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

基本信息

批准号：
8269829
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
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-05-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8269829
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 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

项目摘要

Project Summary 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.

项目概要人们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。这脂质双层阵列的使用有可能提供廉价且高效的高性能吞吐量分析方法能够解决这些问题。有几个关键如果脂质微阵列要取得成功，需要解决的问题通过DNA微阵列分析获得。其中最主要的是需要一个检测脂质微阵列表面药物关联的非侵入性方法由于药物相互作用而导致脂质基质的后续扰动。非线性紫外-可见和频产生（UV-Vis SFG）和红外- 可见光和频发生（IR-Vis SFG）成像可能会给出这个问题的答案问题。非线性成像的几个属性使其成为一种有前途的方法检测微阵列上的药物-膜相互作用，包括量化药物-膜相互作用的能力检测到的信号、高光学分辨率和固有的表面特异性。这实施 UV-Vis SFG 和 IR-Vis SFG 成像的关键第一步脂质微阵列药物关联研究已经实现。具体目标#1 将重点开发用于低浓度检测的 UV-Vis SFG 平面支持的脂质双层（PSLB）中的分子量分子。使用UV- Vis SFG 用于测量蛋白质对膜的吸附，它是上一个资助期的目标也将得到实现。非线性谱将尝试研究几种模型蛋白质和药物靶点的反应更全面地了解目标 #1 中控制 UV-Vis SFG 检测的因素。 UV-Vis SFG 在筛选潜在离子通道抑制剂中的应用在目标#2 中进行了探索。检查药物对热致相的影响通过 IR-Vis SFG 光谱和 IR- 目标 #3 探索了 Vis SFG 高通量成像。这些研究旨在展示非线性成像方法与微图案结合的实用性用于高通量药物筛选应用的流体脂质双层阵列。