Utrasensitive bioassays using palladium catalyzed chemical amplification

使用钯催化化学放大的超灵敏生物测定

• 批准号: 8327721
• 负责人: Mark Lelental
• 金额: $ 10.06万
• 依托单位: BIOFORMATIX, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-02 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8327721
• 关键词: Antibodies; Antigens; Biological Assay; Biological Markers; Biological Models; Blood; Body Fluids; Buffers; CA-125 Antigen; COL1A1 gene; Calibration; Cancer Detection; Cancer Diagnostics; Cancer Patient; Chemicals; Chemistry; Clinical; Complex; Containment; Detection; Development; Diagnostic; Diagnostic Neoplasm Staging; Disease; Dyes; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Equipment; Extinction (Psychology); Formazans; Fred Hutchinson Cancer Research Center; Generic Drugs; Health Care Costs; Human; Immunoassay; Label; Literature; MSLN gene; Maintenance; Malignant neoplasm of ovary; Medical; Metals; Methodology; Methods; Modeling; Molecular; Monitor; Monoclonal Antibodies; Optics; Oxidation-Reduction; Palladium; Patients; Performance; Phase; Procedures; Reaction; Reagent; SLPI gene; Sampling; Screening for cancer; Screening procedure; Serum; Signal Transduction; Specimen; Staging; Survival Rate; Technology; Temperature; Testing; Tetrazolium; Time; Treatment Efficacy; Tumor Antigens; Tumor Markers; Tumor stage; Water; Work; aqueous; assay development; cancer diagnosis; cancer therapy; catalyst; clinical application; density; design; improved; in vitro Assay; malignant breast neoplasm; novel; technology development; tumor; tumor progression

DESCRIPTION (provided by applicant): We propose to use novel Palladium Catalyzed Chemical Amplification (PdCCA) technology to develop ultra-sensitive bioassays that can detect very low concentrations of tumor biomarkers in blood. PdCCA is a generic technology that can be universally applied to different assay platforms. In the bioassay application considered here it uses a detection antibody labeled with a palladium catalyst and a subsequent, palladium catalyzed, high gain redox reaction generating a water-soluble formazan dye. PdCCA will be adapted to the standard ELISA format, where it may provide a factor of 102 to 103 fold increase in sensitivity in immunoassays for which a detection antibody of high selectivity is available. This increase in sensitivity will lower the detection threshold of currently used methods to the desired range of ng-pg/ml, allowing for development of screening biomarkers for early tumor detection. Phase I of the proposed work will concentrate on technology development and optimization using a set of three biomarkers for ovarian cancer and, optionally, one biomarker of breast cancer, currently under study at the Fred Hutchinson Cancer Research Center, Seattle, WA (FHCRC). We will optimize the dye forming amplification chemistry and spectrophotometric detection, and use model in vitro assays to develop and optimize the bioassay technology. Feasibility of our approach will be demonstrated on clinical specimens stored at the FHCRC.

描述(由申请人提供)：我们建议使用新颖的钯催化化学放大(PdCCA)技术来开发能够检测血液中极低浓度的肿瘤生物标志物的超灵敏生物检测方法。PdCCA是一种通用技术，可普遍适用于不同的检测平台。在这里考虑的生物测定应用中，它使用用钯催化剂标记的检测抗体和随后的钯催化的高增益氧化还原反应产生水溶性福尔马赞染料。PdCCA将适应标准的ELISA格式，在具有高选择性检测抗体的免疫分析中，它可以将灵敏度提高102至103倍。这种灵敏度的提高将把目前使用的方法的检测阈值降低到所需的ng-pg/ml范围，从而允许开发用于早期肿瘤检测的筛选生物标记物。拟议工作的第一阶段将集中于技术开发和优化，使用一套卵巢癌的三个生物标记物，以及可选的一个乳腺癌的生物标记物，目前正在华盛顿州西雅图的弗雷德·哈钦森癌症研究中心(FHCRC)进行研究。我们将优化染料形成放大化学和分光光度检测，并利用体外模型检测来开发和优化生物检测技术。我们方法的可行性将在FHCRC储存的临床标本上得到验证。