Nano-scale detection of the disease proteome

疾病蛋白质组的纳米级检测

• 批准号: 7291116
• 负责人: NICHOLAS J WANG
• 金额: $ 12.93万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-24 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291116
• 关键词: Affect; Alternative Splicing; Antibodies; Automation; Basic Science; Binding; Biological Assay; Biological Markers; Biological Models; Biopsy Specimen; Blood; Blood capillaries; Breast; Breast Cancer Cell; Breast Cancer Early Detection; CD44 gene; Cancer cell line; Cells; Chemistry; Classification; Clinical; Color; Compatible; Conditioned Culture Media; Coupled; Cultured Cells; Detection; Development; Devices; Diagnostic; Disease; Early Diagnosis; Engineering; Enzyme-Linked Immunosorbent Assay; Exploratory/Developmental Grant; Fluorescence; Funding; Goals; Immunoassay; Isoelectric Focusing; Knowledge; Laboratories; Light; Malignant Neoplasms; Mammary Neoplasms; Measurement; Membrane; Methods; Modeling; Molecular; Nanotechnology; Nature; Patients; Performance; Phase; Phosphorylation; Post-Translational Protein Processing; Preparation; Procedures; Process; Property; Protein Analysis; Protein Binding; Protein Isoforms; Proteins; Proteolysis; Proteome; Proteomics; Protocols documentation; Quantum Dots; RNA Splicing; Range; Reagent; Reproducibility; Research; Research Personnel; Sampling; Serum; Source; Standards of Weights and Measures; System; Technology; Temperature; Testing; Tumor Cell Line; Variant; Work; capillary; detector; fluorophore; glycosylation; human tissue; improved; malignant breast neoplasm; multidisciplinary; nano; nanoscale; new technology; prognostic; protocol development

DESCRIPTION (provided by applicant): We propose to work collaboratively with investigators funded through this R21 mechanism to develop a new protein detection technology, the nano-capillary immunoassay. This technology will result in significant advances in protein detection and analysis with utility for both basic research and as a clinical diagnostic device. Our group, consisting of researchers at Lawrence Berkeley National Laboratory and Cell Biosciences, Inc., has the multidisciplinary expertise that this project requires. We will use CD44 in breast cancer as our model system to develop and optimize the nano-capillary immunoassay to reach optimal sensitivity (fg/¿L), accuracy, and reproducibility. CD44 is an ideal model protein that will, if we are successful, demonstrate the potential of the technology to identify multiple splice variants and post-translational modifications (e.g., glycosylation, phosphorylation, and proteolysis) in cancer cell lines, tumor samples, and blood serum. Through this funding we will engineer improvements in protein capture chemistry, analyte detection, background reduction, and temperature control to increase sensitivity. We will also develop a new detection system to enable multiplexed analysis of several analytes in a single assay. Standardized protocols will be developed for optimal protein extraction from a range of biologic samples coupled with full automation resulting in high throughput and reproducibility. These approaches will yield a systematic and quantitative approach to protein analysis. This technology fulfills multiple goals of the RFA. 1) The nano-capillary assay offers unbiased detection of protein isoforms and post-translational modifications using a single detection reagent that will produce more complete protein profiles than can be achieved with standard protocols. 2) It will advance our knowledge of the molecular nature of cancer and provide a platform for the discovery of new biomarkers. 3) It has the potential for identifying undiscovered forms of existing biomarkers that may enhance their application as clinical diagnostics. The ultimate goal is to develop a technology to propel cancer biomarker discovery and enable early detection of breast cancer.

描述（由应用程序提供）：我们建议与通过此R21机制资助的调查人员合作，以开发一种新的蛋白质检测技术，即纳米毛细管免疫测定。这项技术将在基础研究和作为临床诊断装置方面的实用性，在蛋白质检测和分析方面取得重大进展。我们的小组由劳伦斯·伯克利国家实验室和细胞生物科学公司的研究人员组成，具有该项目需要的多学科专业知识。我们将在乳腺癌中使用CD44作为模型系统，以开发和优化纳米毛细血管免疫测定，以达到最佳灵敏度（FG/€），准确性和可重复性。 CD44是一种理想的模型蛋白，如果我们成功，它将证明该技术在癌细胞系，肿瘤样品和血清中识别多种剪接变异和翻译后修饰（例如糖基化，磷酸化和蛋白水解）的潜力。通过这项资金，我们将改善蛋白质捕获化学，分析物检测，背景降低和温度控制，以提高灵敏度。 We will also develop a new detection system to enable multiplexed analysis of several analytes in a single assay.将开发标准化方案，以从一系列生物样品中提取最佳蛋白质，并结合完整的自动化，从而产生高吞吐量和可重复性。这些方法将产生一种系统的定量方法来进行蛋白质分析。该技术实现了RFA的多个目标。 1）使用单个检测试剂，纳米毛细血管评估提供了对蛋白质同工型和翻译后修饰的无偏检测，该试剂将产生比使用标准方案可以实现的更完整的蛋白质曲线。 2）它将提高我们对癌症分子性质的了解，并为发现新的生物标志物提供一个平台。 3）它有可能识别未发现的现有生物标志物的形式，这些形式可能会增强其作为临床诊断的应用。最终目标是开发一种技术来推动癌症生物标志物的发现并能够尽早发现乳腺癌。