A Microfluidic Protein Separation Device Based on Dielectrophoresis

一种基于介电泳的微流控蛋白质分离装置

• 批准号: 8314038
• 负责人: Alexandra Ros
• 金额: $ 17.96万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8314038
• 关键词: Address; Adsorption; Alzheimer' s Disease; Amyloid beta-Protein; Automobile Driving; Behavior; Biological Assay; Biomedical Research; Body Fluids; Cerebrospinal Fluid; Charge; Clinical; Complement; Complex; Complex Mixtures; Computer Simulation; Coupled; Detection; Development; Device or Instrument Development; Devices; Diagnosis; Disease Marker; Elements; Gel; Goals; Immunoglobulin Fragments; Injection of therapeutic agent; Investigation; Ionic Strengths; Laboratories; Laws; Malignant - descriptor; Measurement; Methods; Microfluidic Microchips; Microfluidics; Modeling; Molecular Conformation; Molecular Weight; Motion; Nature; Operative Surgical Procedures; Pathway interactions; Peptides; Periodicity; Protein Analysis; Proteins; Recovery; Sample Size; Sampling; Shapes; Site; Solutions; Structure; Surface; System; Techniques; Testing; Time; Variant; base; density; design; electric field; immunoreaction; insight; interest; micro-total analysis system; novel; novel diagnostics; point-of-care diagnostics; prevent; public health relevance; rapid diagnosis; response; single cell analysis; tool

DESCRIPTION (provided by applicant): Reliable and rapid separation of proteins is both a fundamental and challenging problem for bioanalytical and biomedical research. Conventional separation techniques reach their limits at extremes, for example, when increased sample complexity demands for the analysis of relevant disease markers in extremely small concentration and within a huge background. Problems further arise for time critical samples, i.e. when rapid answers are required, such as for samples with temporally degrading or altering composition or when rapid diagnosis is essential, such as during surgery. These limitations can be more drastic for smaller sample volumes and concomitantly low amount of proteins, such as in the case of minimal invasive diagnosis or single cell analysis. The latter is specifically important for understanding specific cellular pathways and malignant progressions, which would otherwise be averaged in the ensemble measurement. Another extremely relevant example represents the diagnosis of Alzheimer's Disease, which is particularly challenging due to the transient nature of the involved peptide species and their extremely low abundance in body fluids. This exploratory proposal aims to develop dielectrophoretic devices for the efficient, rapid and gel-free separation, purification and pre-concentration of proteins on microfluidic platforms. This project applies a new principle for the gel-free separation of proteins in microfluidic systems. It exploits dielectrophoresis (DEP) of proteins, which as polarizable objects respond to a non-uniform electric field with a migrational motion. As the polarizability depends on various parameters such as shape, charge, charge density, permittivity or deformability it thus allows probing the DEP response of proteins in a broad range. The dielectrophoretic response of proteins is provoked in tailored microstructure designs on a lab-on- a-chip platform in which the necessary inhomogeneous electric field gradient can be optimally generated. A fundamental investigation of the DEP behavior of selected proteins will give insight into the necessary electrical driving parameters and reveal optimized conditions for more complex separation problems as well as the purification and pre-concentration of proteins and peptides. This novel device is capable of analyzing protein samples in time scales of a few minutes and reduces sample volumes to the pL-nL range. In particular, we develop a combined microfluidic immunoaffinity and DEP based separation assay for beta-amyloid (A2) oligomers in cerebrospinal fluid. This novel DEP separation method thus represents a development of outstanding importance for biomedical research and point of care diagnostics. PUBLIC HEALTH RELEVANCE (provided by the applicant): The goal of this project is to design and test devices for fast, reliable and gel-free protein separation and pre-concentration based on dielectrophoresis on a microfluidic platform. The development of these devices extends current separation techniques at their limits, i.e. for low sample concentration, time critical analyzes, miniature sample amounts and complex samples. It will facilitate biomedical research and eventually clinical laboratory practice, in particular, rapid and on-site point of care diagnostics.

描述（由申请人提供）：蛋白质的可靠和快速分离既是生物分析和生物医学研究的基本和具有挑战性的问题。例如，当样本复杂性增加对相关疾病标志物的分析以极小的浓度和巨大背景的分析时，传统的分离技术达到了极端的限制。时间临界样本进一步出现问题，即需要快速答案（例如，在时间上降解或改变组成或快速诊断是必不可少的，例如手术期间的样本。对于较小的样品量和及时较低量的蛋白质，例如在最小的侵入性诊断或单细胞分析的情况下，这些局限性可能会更加剧烈。后者对于理解特定的细胞途径和恶性渐进非常重要，否则在整体测量中将平均。另一个极其相关的例子代表了对阿尔茨海默氏病的诊断，由于所涉及的肽物种的短暂性质及其在体液中的极低丰度，这尤其具有挑战性。该探索性提案旨在开发介电性设备，以实现微流体平台上蛋白质的有效，快速和无凝胶分离，纯化和预浓缩。该项目应用了一个新的原理，用于微流体系统中蛋白质无凝胶分离。它利用蛋白质的介电性（DEP），因为可极化物体对具有迁移运动的不均匀电场反应。由于极化性取决于各种参数，例如形状，电荷，电荷密度，介电常数或变形性，因此允许探测蛋白质在广泛范围内的DEP响应。蛋白质的介电性反应在实验室芯片平台上量身定制的微观结构设计中引起，其中可以最佳地生成必要的不均匀电场梯度。对选定蛋白质的DEP行为的基本研究将使您深入了解必要的电驱动参数，并揭示优化条件，以实现更复杂的分离问题以及蛋白质和肽的纯化和预浓缩。这种新型设备能够在几分钟的时间尺度上分析蛋白质样品，并将样品体积减少到PL-NL范围。特别是，我们为脑脊液中的β-淀粉样蛋白（A2）低聚物开发了合并的微流体免疫植入率和基于DEP的分离测定法。因此，这种新型的DEP分离方法代表了对生物医学研究和护理诊断点的杰出重要性的发展。 公共卫生相关性（由申请人提供）：该项目的目的是设计和测试设备，以基于微流体平台上的介电粒细胞性，以快速，可靠和无凝胶的蛋白质分离和预浓缩。这些设备的开发扩展了当前的分离技术，即对于低样本浓度，临界分析，微型样品量和复杂样品。它将促进生物医学研究，并最终促进临床实验室实践，尤其是快速和现场的护理诊断点。