Multiplexed Nanosensing for DNA and Proteins

DNA 和蛋白质的多重纳米传感

• 批准号: 7365327
• 负责人: RICHARD JAMES COTE
• 金额: $ 61.39万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7365327
• 关键词: Address; Antibodies; Base Sequence; Binding; Binding Proteins; Biological Assay; Biological Markers; Biological Models; Biosensing Techniques; Biosensor; Body Fluids; Buffers; CA-125 Antigen; CDKN2A gene; Cancer Patient; Carbon; Carbon Nanotubes; Cell Line; Cells; Characteristics; Chemistry; Clinical; Collaborations; Complex Mixtures; DNA; DNA Methylation; DNA Sequence; DNA lesion; Data; Detection; Development; Devices; Dimensions; Disadvantaged; Disease; Early Diagnosis; Electrolytes; Engineering; Ensure; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Evaluation; Fluorescent Dyes; Gene Mutation; Genes; Genetic; Genetic Polymorphism; Genomics; Goals; HCT116 Cells; Human; Individual; Injection of therapeutic agent; Intervention; Label; Laboratories; Lead; Lesion; Ligands; Low-Density Lipoproteins; Lung; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of urinary bladder; Measurable; Measures; Mediating; Methodology; Methods; Methylation; Microfluidic Microchips; Microfluidics; Modeling; Molecular; Molecular Diagnosis; Monitor; Mutate; Mutation; Nanotubes; Nucleic Acids; Nucleic acid sequencing; Numbers; Oligonucleotide Probes; Oligonucleotides; One-Step dentin bonding system; Operative Surgical Procedures; Optics; Organism; Pathway interactions; Patients; Pattern; Performance; Physiological; Polymerase Chain Reaction; Positioning Attribute; Procedures; Process; Prostate; Prostate-Specific Antigen; Proteins; Public Health; Range; Reaction; Reaction Time; Reading; Recording of previous events; Relative (related person); Reproducibility; Research; Research Personnel; Route; Sampling; Screening for cancer; Semiconductors; Sensitivity and Specificity; Serum; Serum Proteins; Side; Signal Transduction; Silicon; Simulate; Site; Solid; Solutions; Speed; Surface; System; Systems Integration; Technology; Testing; Therapeutic; Transistors; Treatment Efficacy; Validation; Variant; Work; aptamer; base; biochip; bisulfite; cancer diagnosis; cancer type; clinically relevant; cost; cost effective; design; endonuclease; indium oxide; innovation; interest; internal control; mutant; nanobiosensor; nanoscale; nanosensors; nanowire; novel; novel strategies; prognostic; programs; research study; sensor; single walled carbon nanotube; size; success; surface coating

DESCRIPTION (provided by applicant): This application builds on the advances that the USC investigators have made in nanoscale materials and devices, including nanoscale field effect transistors (FETs), microfluidics and biosensors. It has become increasingly clear that it is critical to develop technology platforms that permit quantitative analysis of multiple serum protein biomarkers as well as multiple genetic and epigenetic lesions in a cost-effective manner for early detection, prognostic evaluation and therapeutic monitoring of cancer. To address this critical public health issue, the overall goal of this project is to develop and validate novel nanowire (NW) and single-walled carbon nanotube (SWNT) sensor arrays for single-step detection of serum protein markers, and of DNA mutation, polymorphism and site-specific methylation. This application is focused on the development of multiplexed nanosensor arrays that can be used in a clinical setting. We have demonstrated that In2O3 NW and SWNT- based FETs can be used as effective biosensors with a potential to achieve both high specificity and sensitivity. We propose to develop and test the applicability of the NW/NT FET arrays for multiplexed sensing in model systems as well as in clinical serum samples. Unique surface functionalization for various capture ligands will be achieved via our proven microfluidic delivery system, and semiconductor processing will be employed to create the proposed biosensors. In Specific Aims 1 to 3, we will used novel NW/SWNT sensors to detect 1) cancer related protein biomarkers (including PSA, CEA, CA-125 and other markers as the model); 2) DNA nucleic acid sequence changes related to cancer (using K-ras mutations as the model); and 3) site- specific DNA methylation (epigenetic) cancer related changes (using p16 gene as the model). In Specific Aims 4 and 5, we will integrate a novel microfluidic delivery and test system and develop nanosensors arrays capable of interrogating multiple biomarkers simultaneously. Both NW and SWNT-based devices will be integrated side-by-side into the same array, a powerful design which will allow complementarily of signals for analyte binding, minimizing false negative or positive signals for complex mixtures. The sensitivity, selectivity, stability and reproducibility of these sensor arrays will be investigated. In Specific Aim 6, we will test serum from cancer patients with known levels of relevant biomarkers. The applications addressed here will result in sensor array platforms that are highly likely to yield label-free, low-cost, multiplexed assays for molecular diagnosis and monitoring therapeutic efficacy for cancer, and which can easily be extended to applications to other diseases as well. Project Narrative/Relevance: It is becoming increasingly clear that in order to achieve the goal of early detection and therapeutic monitoring of cancer using serum based biomarkers, including proteins, DNA mutations, and other changes in DNA, it is critical to develop technology platforms that permit quantitative analysis of multiple biomarkers simultaneously and a cost effective manner. In order to address this critical public health issue, the overall goal of this project is to develop and validate novel nanowire (NW) and single-walled carbon nanotube (SWNT) sensor arrays for single step detection of serum based protein and DNA biomarkers. The research in this application is highly likely to result in novel devices that will allow for the detection of multiple cancer associated biomarkers capable of use in early detection and therapeutic monitoring of cancer. It is also likely that this device will be easily extended to applications in other diseases as well.

描述（由申请人提供）：此申请基于USC研究人员在纳米级材料和设备中所做的进步，包括纳米级现场效应晶体管（FET），微流体和生物传感器。越来越清楚的是，开发技术平台至关重要，以允许对多种血清蛋白生物标志物以及多种遗传和表观遗传病变进行定量分析，以具有成本效益的方式进行早期检测，预后评估和对癌症的治疗监测。为了解决这个关键的公共卫生问题，该项目的总体目标是开发和验证新颖的纳米线（NW）和单壁碳纳米管（SWNT）传感器阵列，用于单步检测血清蛋白质标记，以及DNA突变，多态性和特定于特定的甲基化。该应用的重点是可以在临床环境中使用的多路复用纳米传感器阵列的开发。我们已经证明，In2O3 NW和SWNT基于基于SWNT的FET可以用作有效的生物传感器，具有具有高特异性和敏感性的潜力。我们建议在模型系统以及临床血清样品中开发和测试NW/NT FET阵列在多重传感中的适用性。将通过我们经过验证的微流体递送系统实现各种捕获配体的独特表面功能，并将采用半导体加工来创建所提出的生物传感器。在特定的目标1到3中，我们将使用新颖的NW/SWNT传感器检测1）癌症相关的蛋白质生物标志物（包括PSA，CEA，CA-125和其他标记物作为模型）； 2）与癌症有关的DNA核酸序列变化（使用K-RAS突变作为模型）； 3）位点特异性DNA甲基化（表观遗传）癌症相关的变化（使用p16基因作为模型）。在特定的目标4和5中，我们将整合一种新型的微流体传递和测试系统，并开发能够同时询问多个生物标志物的纳米传感器阵列。基于NW和SWNT的设备都将并排集成到同一阵列中，这是一种强大的设计，可以互补地进行分析物结合的信号，从而最大程度地减少复杂混合物的假阴性或正信号。这些传感器阵列的灵敏度，选择性，稳定性和可重复性将被研究。在特定目标6中，我们将测试具有已知水平相关生物标志物的癌症患者的血清。这里解决的应用将导致传感器阵列平台，这些平台很可能产生无标签，低成本，多重分析的测定法，以用于分子诊断和监测癌症的治疗功效，并且很容易将其扩展到其他疾病的应用。 项目叙事/相关性：越来越清楚的是，要使用基于血清的生物标志物（包括蛋白质，DNA突变和其他DNA变化）来实现对癌症进行早期检测和治疗性监测的目标，这对于开发技术平台的至关重要的是允许对多个生物标记物进行定量分析的技术平台至关重要。为了解决这个关键的公共卫生问题，该项目的总体目标是开发和验证新颖的纳米线（NW）和单壁碳纳米管（SWNT）传感器阵列，以单步检测基于血清的蛋白质和DNA生物标志物。该应用中的研究极有可能导致新型设备，从而可以检测能够在癌症早期检测和治疗监测中使用的多种癌症相关的生物标志物。该设备也可能很容易扩展到其他疾病的应用。