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首页> 外文学位 >Towards early stage disease detection in microdevices: Fabrication and testing of micro total analysis systems for bioanalytical applications.
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Towards early stage disease detection in microdevices: Fabrication and testing of micro total analysis systems for bioanalytical applications.

机译：面向微型设备的早期疾病检测：用于生物分析应用的微型整体分析系统的制造和测试。

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摘要

The past few years have seen a rapid expansion in interest in the characterization of the entire complement of proteins, or proteome. Micro total analysis systems (muTAS) are an emerging promising method, offering rapid, sensitive and low sample consumption separations.;I have also shown the evaluation of poly(methylmethacrylate) (PMMA) and thermoset polyester (TPE) microchips for use in protein profiling. To improve separation efficiency and reduce protein adsorption, dynamic coating and poly(ethylene glycol) (PEG) grafting using atom transfer radical polymerization (ATRP) have been used in PMMA microdevices. Proteins, peptides and protein digests have been separated electrophoretically in these PMMA microchips. My results demonstrate that PMMA microdevices should be well suited as microfluidic systems for high performance separations of complex biological mixtures.;In-channel ATRP has been developed for the surface modification of TPE microdevices. Characterization indicates that PEG-modified microchannels have much lower and more pH-stable electroosmotic flow, more hydrophilic surfaces and reduced nonspecific protein adsorption. CE of amino acid and peptide mixtures in these PEG-modified TPE microchips had good reproducibility. Phosducin-like protein and phosphorylated phosducin-like protein were also separated to measure the phosphorylation efficiency. My results show that PEG-grafted TPE microchips have broad potential application in biomolecular analysis.;Cancer marker analysis is important for medical research and applications. I report a method that can covalently attach appropriately oriented antibodies of interest on monolith surfaces. To reduce nonspecific adsorption, protein solutions were used to effectively block the monolith surface. Selective preconcentration and elution of human chorionic gonadotropin have been performed in my affinity columns, demonstrating that this type of system should have promising applications in cancer marker detection.;I have demonstrated microchip capillary electrophoresis (CE) devices made of CaF2. New methods have been developed for micromachining enclosed capillaries in CaF2. CE analysis of fluorescently labeled amino acids was used to illustrate bioanalytical applications of these microdevices. Initial on-chip infrared spectroscopy results for qualitative analyte identification were achieved in microfluidic CaF2 channels.
机译：在过去的几年中，人们对蛋白质或蛋白质组的整个补充特征的兴趣迅速增长。微量总分析系统（muTAS）是一种新兴的有前途的方法，提供了快速，灵敏和低样品消耗的分离方法;我还展示了对用于蛋白质谱分析的聚甲基丙烯酸甲酯（PMMA）和热固性聚酯（TPE）微芯片的评估。为了提高分离效率并减少蛋白质吸附，在PMMA微型设备中使用了原子转移自由基聚合（ATRP）的动态涂层和聚（乙二醇）（PEG）接枝技术。在这些PMMA微芯片中，蛋白质，肽和蛋白质消化物已通过电泳分离。我的结果表明，PMMA微型设备应非常适合用作微流体系统，以高效分离复杂的生物混合物。;通道内ATRP已开发用于TPE微型设备的表面改性。表征表明，PEG修饰的微通道具有更低的pH稳定性和更高的电渗流量，更多的亲水表面以及减少的非特异性蛋白质吸附。这些PEG修饰的TPE微芯片中氨基酸和肽混合物的CE具有良好的重现性。还分离出类似磷光诱导素的蛋白和磷酸化的磷光诱导素样蛋白，以测量磷酸化效率。我的结果表明，PEG接枝的TPE芯片在生物分子分析中具有广泛的应用潜力。癌症标志物分析对医学研究和应用具有重要意义。我报告了一种方法，该方法可以将适当定向的目标抗体共价结合在整料表面上。为了减少非特异性吸附，蛋白质溶液被用来有效地阻塞整体表面。在我的亲和柱中已经进行了人绒毛膜促性腺激素的选择性预浓缩和洗脱，表明这种类型的系统在癌症标志物检测中应具有广阔的应用前景。我已经证明了由CaF2制成的微芯片毛细管电泳（CE）装置。已经开发出用于微加工CaF2中封闭毛细管的新方法。荧光标记氨基酸的CE分析用于说明这些微型设备的生物分析应用。初步的片上红外光谱结果用于定性分析物鉴定是在微流体CaF2通道中实现的。

著录项

  • 作者

    Pan, Tao.;

  • 作者单位

    Brigham Young University.;

  • 授予单位 Brigham Young University.;
  • 学科 Biochemistry.
  • 学位 Ph.D.
  • 年度 2007
  • 页码 152 p.
  • 总页数 152
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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