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(申请人提供)：拟议研究的目标是开发可靠和广泛适用的方法，用于在微升样本量中以极低的(毫微摩尔)浓度进行蛋白质生物标志物的多重体外检测。这些新颖的超灵敏微阵列生物传感测量将通过结合使用表面酶化学、生物功能化纳米颗粒和独特的基于表面等离子体的生物亲和吸附检测技术来实现。这些新方法的关键部分是(I)使用酶反应在纳米颗粒表面和微阵列元件上连接、修饰和放大生物分子，以提高生物亲和力传感过程的选择性和灵敏度；(Ii)将化学和酶生物功能化的纳米颗粒纳入微阵列传感测量中，以增强对目标生物分子的捕获和产生的信号；以及(Iii)实施新颖的光学检测方法，如表面等离子体共振相位成像和纳米颗粒增强的光学衍射，以提供多路复用形式的额外生物传感灵敏度。适体、抗体和酶附着反应的组合将被用来捕获和检测存在于齐妥莫水平的生物流体样品中的蛋白质(例如，血管内皮生长因子、PSA、肌钙蛋白)和微型RNA生物标志物靶标。这些新的检测方法可以识别和量化飞摩尔浓度及以下的多种蛋白质，这是发现新的蛋白质生物标记物集合的关键技术，能够在如此低的水平跟踪这些新的生物标记物(例如，心血管和癌症生物标记物)，将有助于创建更好的早期疾病检测方法和治疗后患者监测策略。