Bioaffinity Assays Using UV One-Dimensional Photonic Crystals (1DPC)

使用紫外一维光子晶体 (1DPC) 进行生物亲和力测定

• 批准号: 8957305
• 负责人: Joseph R. LAKOWICZ
• 金额: $ 22.74万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8957305
• 关键词: Affect; Albumins; Antibodies; Area; Basic Science; Binding; Binding Proteins; Biological; Biological Assay; Biological Models; Biological Testing; Biomedical Research; Bovine Serum Albumin; Bypass; CD4 Antigens; Capsid Proteins; Chemicals; Collaborations; Complex; Coupled; Coupling; DC-specific ICAM-3 grabbing nonintegrin; Deposition; Detection; Devices; Dimensions; Dyes; Film; Fluorescence; Future; Generations; Generic Drugs; HIV; HIV Antibodies; HIV Envelope Protein gp120; Immunoglobulin G; Kinetics; Label; Lighting; Link; MDM2 gene; Measurement; Measures; Metals; Methods; Modeling; Optics; Performance; Plasma; Protein Array; Protein Binding; Protein p53; Proteins; Proteomics; Radiation; Reaction; Reader; Refractive Indices; Reporting; Research; Sampling; Signal Transduction; Silicon Dioxide; Spatial Distribution; Streptavidin; Structure; Surface; Surface Plasmon Resonance; System; Tantalum; Temperature; Testing; Tryptophan; Tumor Suppressor Proteins; Utah; Work; anti-IgG; base; biological systems; cost; design; drug discovery; fluorophore; instrument; instrumentation; monolayer; novel strategies; photonics; prevent; protein protein interaction; public health relevance; receptor; research clinical testing; screening; sensor; silicon nitride; simulation; transmission process; vapor

 DESCRIPTION (provided by applicant): Measurements of biomolecule interactions are increasingly important for proteomics and protein-protein interactions (the Interactome). With the exception of surface plasmon resonance (SPR) most of the alternative methods rely on the use of extrinsic fluorophores or probes which are covalently linked to one or more of the interacting species. These labeling steps become increasingly costly and complex as the number of biomolecules is increased. We propose a new approach to label-free measurements using one-dimensional photonic crystals (1DPCs) and intrinsic protein fluorescence. Since most proteins contain tryptophan these residues provide a type of mass sensor for proteins, analogous to SPR which also senses mass by changes in the refractive index. We will use Bloch surface waves (BSWs) on the 1DPCs. BSWs provide opportunities for highly localized excitation and observation of surface-bound proteins. The BSW substrates will be composed of multiple layers of UV-transmitting dielectrics. These structures will not contain metals so there is no quenching at short distances. Specific Aim 1. Design BSW structures for use at UV wavelengths. Not all 1DPCs display BSW. We will use the transfer matrix method to simulate the optical modes and select the dielectrics and dimensions. More complex numerical simulations will be used to predict the efficiency of trp coupling and directionality of the emission. Specific Aim 2. Fabricae and test 1DPCs for use with UV wavelengths. Suitable structures must display good UV transmission, a BSW and low autofluorescence. We will test combinations of three dielectrics, silica (SiO2), silicon nitride (Si3N4) and tantalum pentoxide (Ta2O5). Substrates will be tested wih model systems of albumin, streptavidin and antibodies. Specific Aim 3. Biological testing of the BSW structures. The structures will be tested with two important biological systems. The first is binding between the p53 tumor suppressor protein and its negative effectors. The second is binding of the HIV gp120 protein with its CD4 receptor and with potential HIV antibodies. Impact. This project will result in a new generic method to measure protein-protein interactions. The substrates will be simple and inexpensive. The method can be used with existing or modified plate readers. The simplicity of the optics will permit the extension to high-throughput measurements.

 描述（由适用提供）：生物分子相互作用的测量对于蛋白质组学和蛋白质 - 蛋白质相互作用（相互作用组）越来越重要。除表面等离子体共振（SPR）外，大多数替代方法都取决于外在荧光团或与一个或多个相互作用物种共价相关的问题。随着生物分子数量的增加，这些标记步骤变得越来越成本和复杂。我们提出了一种使用一维光子晶体（1DPC）和内在蛋白质荧光的新方法来无标记测量。由于大多数蛋白质含有色氨酸，这些残基为蛋白质提供了一种类型的质量传感器，类似于SPR，这也通过折射率的变化来感知质量。我们将在1DPC上使用Bloch表面波（BSW）。 BSW为高度局部的兴奋和观察表面结合蛋白提供了机会。 BSW底物将由多层紫外线传播饮食学组成。这些结构将不包含金属，因此在短距离内没有淬火。特定目标1。设计BSW结构，用于在UV波长下使用。并非所有1DPC显示BSW。我们将使用转移矩阵方法模拟光学模式并选择饮食和尺寸。更复杂的数值模拟将用于预测TRP耦合的效率和方向性特定的目标2。织物和测试1DPC，可与紫外线波长一起使用。合适的结构必须显示出良好的紫外线传输，BSW和低自动荧光。我们将测试三种饮食（SIO2），氮化硅（SI3N4）和坦塔坦五氧化物（TA2O5）的组合。将测试底物的白蛋白，链霉亲和抗体的WIH模型系统。特定目的3。BSW结构的生物测试。这些结构将通过两个重要的生物系统进行测试。第一个是p53肿瘤抑制蛋白及其负面影响之间的结合。第二个是HIV GP120蛋白与其CD4受体和潜在的HIV抗体结合。影响。该项目将导致一种测量蛋白质 - 蛋白质相互作用的新通用方法。底物将是简单且便宜的。该方法可以与现有或修改的板读取器一起使用。光学器件的简单性将允许扩展到高通量测量。