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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：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，除了表面等离子共振（SPR）之外，大多数替代方法都依赖于外源荧光团或探针的使用。随着生物分子数量的增加，这些标记步骤变得越来越昂贵和复杂，我们提出了一种新的无标记测量方法。使用一维光子晶体 (1DPC) 和内在蛋白质荧光，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质量传感器，类似于 SPR，它也通过折射率的变化来感知质量。 1DPC 上的 BSW 为表面结合蛋白的高度局部激发和观察提供了机会。BSW 基底将由多层紫外线传输电介质组成。结构不包含金属，因此在短距离内不会发生猝灭。具体目标 1. 设计用于 UV 波长的 BSW 结构，我们将使用传输矩阵方法来模拟光学模式并选择电介质和材料。更复杂的数值模拟将用于预测色氨酸耦合的效率和发射的方向性。 2. 制造和测试适用于 UV 波长的 1DPC。结构必须表现出良好的紫外线透射率、BSW 和低自发荧光。我们将使用白蛋白、链霉亲和素和五氧化二钽的模型系统来测试三种电介质：二氧化硅 (SiO2)、氮化硅 (Si3N4) 和五氧化二钽 (Ta2O5) 的组合。具体目标 3. BSW 结构的生物学测试该结构将通过两个重要的生物系统进行测试。第二个是 HIV gp120 蛋白与其 CD4 受体和潜在的 HIV 抗体的结合。该项目将产生一种新的通用方法来测量蛋白质-蛋白质相互作用。该方法可以与现有的或改进的读板器一起使用，其光学器件的简单性将允许扩展到高通量测量。