Cancer Sample Preparation with Micromachined Magnetic Sifter and Nanoparticles

使用微机械磁力筛和纳米颗粒制备癌症样品

• 批准号: 8054822
• 负责人: SHAN X. WANG
• 金额: $ 27.37万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8054822
• 关键词: Acute; Address; Affinity; Albumins; Antibodies; Area; Biochemistry; Biological; Biological Assay; Biological Markers; Biosensor; Blood; Blood specimen; Cancer Diagnostics; Cell Culture Techniques; Cell Size; Cell secretion; Cells; Clinical; Clinical Research; Collection; Communities; Complex; Coupled; Cytolysis; Detection; Diagnostic; Dimensions; Exhibits; Film; Fluorescence; Fluorescence-Activated Cell Sorting; Grant; Harvest; Hematologic Neoplasms; Hour; Human; Investigation; Label; Laboratories; Liquid substance; Magnetism; Malignant Neoplasms; Malignant neoplasm of lung; Mass Spectrum Analysis; Methods; Microscopy; Molecular; Molecular Diagnosis; Nanotechnology; Noise; Optics; Outcome; Patients; Performance; Positioning Attribute; Preparation; Procedures; Process; Protein Analysis; Protein Microchips; Proteins; Proteomics; Protocols documentation; Reagent; Reproducibility; Running; Sampling; Sensitivity and Specificity; Serum; Shapes; Signal Transduction; Solid; Solid Neoplasm; Solutions; Speed; Structure; TNF gene; Techniques; Technology; Tube; Validation; Variant; Work; anticancer research; base; biochip; cancer cell; cancer stem cell; cancer therapy; cell injury; cell type; clinical practice; cost; cross reactivity; design and construction; fighting; implantation; improved; innovation; innovative technologies; iron oxide; magnetic field; nano; nanometer; nanoparticle; neoplastic cell; novel; oncology; particle; peripheral blood; quantum; research study; sample collection; tool; tumor

DESCRIPTION (provided by applicant): Variations in sample preparation may contribute to major discrepancies in the quantity and type of biomolecules or cells identified by different laboratories, even though the same reagents and biosensors (or biochips) are employed. This is especially hampering our efforts to fight cancer as sample preparation is a prerequisite for cancer biomarker discovery, validation, and their use in molecular diagnosis and treatment of patients. This urgent need for cancer sample preparation requires innovative methods to bring about better and more affordable sample preparation tools. We have formed an interdisciplinary team with expertise from magnetics, nanotechnology, cancer clinical practice and research, biochemistry, and proteomics to work on a novel micromachined magnetic sifter and directly fabricated magnetic nanoparticles. The magnetic sifter can be used to separate and enrich cancer targets; including proteins and rare cells from raw samples. The utilization of precisely dimensioned structures, including the use of sub-nanometer control of quantum magnetic effects will enable high throughput, high capture yield, low impurities, and low cost. The technology is very unique because high magnetic field gradients (>1 Tesla/ 5m) and large flow rates (>1 mL/hour) are enabled by three-dimensional Si-based micromachining. The use of high-moment magnetic nanoparticles with distinctive shapes and controlled magnetic chain formation give rise to unprecedented capture and release efficiencies, cell damage minimization, enhanced characterization, and run-to-run reproducibility. The proposed project is organized in three aims: Specific Aim 1. Magnetic sifter for high efficiency capture and analysis of protein cancer markers and cancer cells: a) Construct a magnetic sifter with a high magnetic field gradient (>1 Tesla/ 5m) and large flow rate (>1 mL/hour) and demonstrate the efficient capture, release and analysis of protein cancer markers. b) Use magnetic sifter to capture and release NCI-H1650 lung cancer tumor cells cancer cells spiked into samples of human blood with an overall capture efficiency of >70%, while keeping cells viable. Specific Aim 2. Functionalization and characterization of novel magnetic nanoparticles for high efficiency high speed magnetic separation. Specific Aim 3. Magnetic sifter for rare cells capture and analysis from human blood. The utility of the grant includes drastically reducing post-separation analysis and enabling new investigations in areas where rare molecules and cell types are currently too difficult to obtain and analyze. Furthermore, separating protein and cancer markers from high concentration impurities in blood, with 1000-fold enrichment, will enable detection of low abundance cancer markers which are difficult to quantify with conventional technologies. We expect that the tools developed under this grant will have a great impact on the cancer research and treatment community.

描述（由申请人提供）：样品制备的变化可能会导致不同实验室鉴定的生物分子或细胞的主要差异，即使采用了相同的试剂和生物传感器（或生物传感器（或生物芯片））。这尤其阻碍了我们与癌症作斗争的努力，因为样本制备是癌症生物标志物发现，验证及其在分子诊断和治疗患者中的使用的先决条件。对癌症样品制备的这种迫切需求需要创新的方法来提供更好，更负担得起的样本准备工具。我们组成了一个跨学科团队，具有磁性，纳米技术，癌症临床实践和研究，生物化学和蛋白质组学的专业知识，以从事新型的微机械磁性磁带和直接制造的磁性纳米颗粒。磁性筛子可用于分离和富集癌症靶标。包括原始样品中的蛋白质和稀有细胞。精确尺寸结构的利用，包括使用量子磁效应的亚纳米控制，将使高吞吐量，高捕获产量，低杂质和低成本。该技术非常独特，因为高磁场梯度（> 1 Tesla/ 5m）和大型流速（> 1 mL/小时）可以通过三维基于SI的微加工来实现。使用具有独特形状和受控磁链形成的高弹磁性纳米颗粒的使用会导致前所未有的捕获和释放效率，细胞损伤最小化，增强的表征以及运行到运行的可重复性。提出的项目在三个目标中组织：特定目标1。用于高效捕获和分析蛋白质癌标志物和癌细胞的磁性磁带：a）构建具有高磁场梯度（> 1 tesla/ 5m）和大型磁场sifter流速（> 1 mL/小时），并证明了蛋白质癌标志物的有效捕获，释放和分析。 b）使用磁sifter捕获和释放NCI-H1650肺癌肿瘤细胞癌细胞掺入人体血液样本中，总体捕获效率> 70％，同时保持细胞可行。特定目标2。新型磁性纳米颗粒的功能化和表征，用于高效率高速磁分离。特定目标3。稀有细胞的磁性磁纤维捕获和分析人类血液。 该赠款的效用包括大幅度降低分离后分析，并在目前难以获得和分析的罕见分子和细胞类型的领域进行新的研究。此外，将蛋白质和癌症标志物与血液中的高浓度杂质分开，具有1000倍的富集，可以检测到低丰度癌标记物，这些标志物很难用常规技术来量化。我们预计，根据这笔赠款开发的工具将对癌症研究和治疗社区产生重大影响。