Multiplexed Nanosensing for DNA and Proteins

DNA 和蛋白质的多重纳米传感

• 批准号: 7676752
• 负责人: RICHARD JAMES COTE
• 金额: $ 62.08万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 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 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676752
• 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; Oligonucleotide Probes; Oligonucleotides; One-Step dentin bonding system; Operative Surgical Procedures; Optics; Organism; Pathway interactions; Patients; Pattern; Performance; Physiological; Positioning Attribute; Procedures; Process; Prostate; Prostate-Specific Antigen; Proteins; Public Health; 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; success; surface coating; therapeutic evaluation

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）：本申请建立在南加州大学研究人员在纳米级材料和设备方面取得的进展的基础上，包括纳米级场效应晶体管（FET）、微流体和生物传感器。越来越明显的是，开发能够以经济有效的方式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的技术平台对于癌症的早期检测、预后评估和治疗监测至关重要。为了解决这一关键的公共卫生问题，该项目的总体目标是开发和验证新型纳米线 (NW) 和单壁碳纳米管 (SWNT) 传感器阵列，用于单步检测血清蛋白标记物和 DNA 突变，多态性和位点特异性甲基化。该应用的重点是开发可用于临床环境的多路复用纳米传感器阵列。我们已经证明，基于 In2O3 NW 和 SWNT 的 FET 可用作有效的生物传感器，具有实现高特异性和灵敏度的潜力。我们建议开发和测试 NW/NT FET 阵列在模型系统以及临床血清样本中的多重传感的适用性。通过我们经过验证的微流体输送系统将实现各种捕获配体的独特表面功能化，并且将采用半导体加工来创建所提出的生物传感器。在具体目标1至3中，我们将使用新型NW/SWNT传感器来检测1）癌症相关蛋白质生物标志物（包括PSA、CEA、CA-125等标志物作为模型）； 2）与癌症相关的DNA核酸序列变化（以K-ras突变为模型）； 3）位点特异性DNA甲基化（表观遗传）癌症相关变化（使用p16基因作为模型）。在具体目标 4 和 5 中，我们将集成新型微流体输送和测试系统，并开发能够同时询问多个生物标记物的纳米传感器阵列。基于 NW 和 SWNT 的设备将并排集成到同一个阵列中，这是一种强大的设计，可以实现分析物结合信号的互补，最大限度地减少复杂混合物的假阴性或阳性信号。将研究这些传感器阵列的灵敏度、选择性、稳定性和再现性。在具体目标 6 中，我们将测试癌症患者的血清中相关生物标志物水平已知的情况。这里讨论的应用将产生传感器阵列平台，该平台很可能产生无标记、低成本、多重检测，用于分子诊断和监测癌症的治疗效果，并且也可以轻松扩展到其他疾病的应用。 项目叙述/相关性：越来越清楚的是，为了实现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）对癌症进行早期检测和治疗监测的目标，开发能够实现以下目标的技术平台至关重要：允许同时以具有成本效益的方式对多个生物标志物进行定量分析。为了解决这一关键的公共卫生问题，该项目的总体目标是开发和验证新型纳米线 (NW) 和单壁碳纳米管 (SWNT) 传感器阵列，用于单步检测基于血清的蛋白质和 DNA 生物标志物。该应用的研究极有可能产生新型设备，该设备将允许检测多种癌症相关生物标志物，这些生物标志物能够用于癌症的早期检测和治疗监测。该设备也可能很容易扩展到其他疾病的应用。

项目成果

Selective, electrochemically activated biofunctionalization of In2O3 nanowires using an air-stable surface modifier.

使用空气稳定的表面改性剂对 In2O3 纳米线进行选择性电化学激活生物功能化。

• 发表时间: 2011-12
• 影响因子: 9.5
• 作者: Zhang, Rui;Curreli, Marco;Thompson, Mark E
• 通讯作者: Thompson, Mark E

Rapid, label-free, electrical whole blood bioassay based on nanobiosensor systems.

基于纳米生物传感器系统的快速、无标记、电子全血生物测定。

• DOI: 10.1021/nn2035796
• 发表时间: 2011-11-10
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Hsiao;F. Ishikawa;Rui Zhang;R. Datar;R. Cote;M. Thompson;Chongwu Zhou
• 通讯作者: Chongwu Zhou

Highly scalable, uniform, and sensitive biosensors based on top-down indium oxide nanoribbons and electronic enzyme-linked immunosorbent assay.

基于自上而下的氧化铟纳米带和电子酶联免疫吸附测定的高度可扩展、均匀且灵敏的生物传感器。

• DOI: 10.1021/nl5047889
• 发表时间: 2015-02-05
• 期刊: Nano letters
• 影响因子: 10.8
• 作者: N. Aroonyadet;Xiaoli Wang;Yan Song;Haitian Chen;R. Cote;M. Thompson;R. Datar;Chongwu Zhou
• 通讯作者: Chongwu Zhou

A calibration method for nanowire biosensors to suppress device-to-device variation.

一种用于抑制设备间差异的纳米线生物传感器的校准方法。

• DOI: 10.1021/nn9011384
• 发表时间: 2009-12-22
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Ishikawa FN;Curreli M;Chang HK;Chen PC;Zhang R;Cote RJ;Thompson ME;Zhou C
• 通讯作者: Zhou C

Label-free, electrical detection of the SARS virus N-protein with nanowire biosensors utilizing antibody mimics as capture probes.

利用纳米线生物传感器利用抗体模拟物作为捕获探针，对 SARS 病毒 N 蛋白进行无标记电检测。

• DOI: 10.1021/nn900086c
• 发表时间: 2009-05-26
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Ishikawa, Fumiaki N.;Chang, Hsiao-Kang;Curreli, Marco;Liao, Hsiang-I;Olson, C. Anders;Chen, Po-Chiang;Zhang, Rui;Roberts, Richard W.;Sun, Ren;Cote, Richard J.;Thompson, Mark E.;Zhou, Chongwu
• 通讯作者: Zhou, Chongwu