Multiplexed Surface Plasmonic Biomarker Detection with Enzymatic Amplification

通过酶扩增进行多重表面等离子体生物标志物检测

• 批准号: 8517734
• 负责人: ROBERT M CORN
• 金额: $ 29.48万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8517734
• 关键词: Adsorption; Aftercare; Amplifiers; Antibodies; Area; Binding; Biochemical Reaction; Biological; Biological Markers; Biosensing Techniques; Cardiovascular system; Chemistry; Complex; Coupled; DNA; DNA Ligases; DNA-Directed DNA Polymerase; Detection; Development; Disease; Elements; Enzymes; Goals; Gold; Heart Diseases; Image; In Vitro; Knowledge; Label; Ligation; Liquid substance; Magnetism; Malignant Neoplasms; Measurement; Methodology; Methods; MicroRNAs; Modification; N.I.H. Research Support; Nucleic Acids; Optics; Patient Monitoring; Phase; Process; Proteins; RNA; Reaction; Research; Ribonuclease H; Role; Sampling; Scheme; Set protein; Signal Transduction; Silicon Dioxide; Surface; Surface Plasmon Resonance; Techniques; Technology; Troponin; Vascular Endothelial Growth Factors; aptamer; base; detector; ferrite; imaging modality; improved; monolayer; nanoparticle; nanorod; novel; plasmonics

DESCRIPTION (provided by applicant): The goal of the proposed research is the development of reliable and widely applicable methods for the multiplexed in vitro detection of protein biomarkers at extremely low (femtomolar) concentrations in microliter sample volumes. These novel ultrasensitive microarray biosensing measurements will be achieved through the combined use of surface enzyme chemistries, biofunctionalized nanoparticles, and unique surface plasmon-based bioaffinity adsorption detection techniques. Key components in these new methodologies are (i) the use of enzymatic reactions for the attachment, modification and amplification of biomolecules on both nanoparticle surfaces and microarray elements to increase the selectivity and sensitivity of the bioaffinity sensing process, (ii) the incorporation of chemically and enzymatically biofunctionalized nanoparticles into the microarray sensing measurements in order to enhance the capture of and signal generated by target biomolecules, and (iii) the implementation of novel optical detection methods such as surface plasmon resonance phase imaging and nanoparticle-enhanced optical diffraction that provide additional biosensing sensitivity in a multiplexed format. A combination of aptamers, antibodies and enzymatic attachment reactions will be used to capture and detect protein (e.g., VEGF, PSA, troponin) and microRNA biomarker targets that exist in biofluid samples at zeptomole levels. These new detection methodologies that can identify and quantitate multiple proteins at femtomolar concentrations and below are essential technologies for the discovery of new sets of protein biomarkers, and the ability to track these new biomarkers (e.g., cardiovascular and cancer biomarkers) at such low levels will facilitate the creation of better methods for early disease detection and strategies for post-treatment patient monitoring.

描述（由申请人提供）：拟议的研究的目的是开发可靠且广泛适用的方法，用于在微氧样品量中极低（femtomall）浓度的蛋白质生物标志物在体外检测。这些新型的超敏化微阵列生物传感测量将通过表面酶化学，生物功能化的纳米颗粒和独特的基于表面等离子体的基于表面的生物影响力吸附检测技术来实现。这些新方法中的关键组成部分是（i）使用酶反应在纳米颗粒表面和微阵列元素上的依恋，修改和放大生物分子的依恋，修改和扩增，以提高生物伴侣感应过程的选择性和敏感性，（ii）在化学和辅助型物质中逐步启动型号，以启动型号的型号，并逐渐启动nanopartiles Incortiles Incortiles Incortiles Incortiles Incortiles Incortiles Incortiles Incortiles Incoriates Incortiles Incoriates Incortiple inanoptiption inanoptiption inanoptipt目标生物分子产生的捕获和信号，以及（iii）新型光学检测方法的实施，例如表面等离子体共振相成像和纳米颗粒增强的光学衍射，以多种形式提供额外的生物敏感性。适体，抗体和酶促附着反应的组合将用于捕获和检测蛋白质（例如VEGF，PSA，Troponin）和MicroRNA生物标志物靶标，这些靶标在生物氟样品中存在于Zeptomole水平。这些新的检测方法可以识别和定量在女权摩尔浓度下及以下的多种蛋白质，这是发现新的蛋白质生物标志物集合的必要技术，并且能够跟踪这些新的生物标志物（例如，在这种低水平的心血管血管和癌症生物标志物）上以这种较低的水平来促进更好的早期疾病探测和策略的方法。