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Development Of Microfluidic Devices For Biopharmaceutical Production And Biotoxin Detection

机译:用于生物制药和生物毒素检测的微流控装置的开发

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摘要

In this work, we present two different applications of microfluidic control. In the first application, we have developed a microfluidic device that has the potential to automate combinatorial protocols like protein refolding, impacting the biopharmaceutical industry. This device uses microfluidic valves and pumps that can be operated in an automated fashion for fluidic control. We performed refolding experiments on the protein [beta]-galactosidase and showed on-chip quantification of refolding yield using a fluorometric assay. In the second application, we have developed a microfluidic immunobiosensor with an integrated preconcentration system to improve the detection sensitivity. A nanoporous membrane fabricated in-situ using photopolymerization technique inside glass microchannels acts as the preconcentration system. Analytes were electrophoretically concentrated at the membrane and the concentrated bolus was eluted towards a detection region downstream. We performed proof-of-principle experiments using biotin-streptavidin binding to show the improvement in detection sensitivity of this device as opposed to a device that does not include a preconcentration system. Using this device, we also showed a detection limit of 1.6 X 105 PFU/ml for Feline Calicivirus (FCV), which is a model system for human enteric virus. This device has potential to serve as an early detection system for such enteric viruses in environmental water samples.
机译:在这项工作中,我们介绍了微流控的两种不同应用。在第一个应用程序中,我们开发了一种微流控设备,它具有使诸如蛋白质复性之类的组合协议自动化的潜力,从而影响了生物制药行业。该设备使用微流体阀和泵,这些阀和泵可以自动方式进行流体控制。我们对蛋白质β-半乳糖苷酶进行了重折叠实验,并使用荧光测定法显示了重折叠产量的芯片上量化。在第二个应用程序中,我们开发了具有集成预浓缩系统的微流体免疫生物传感器,以提高检测灵敏度。在玻璃微通道内部使用光聚合技术原位制造的纳米多孔膜可作为预浓缩系统。将分析物电泳浓缩在膜上,并将浓缩的大丸剂向下游的检测区域洗脱。我们进行了使用生物素-链霉亲和素结合的原理验证实验,以显示与不包含预浓缩系统的设备相比,该设备的检测灵敏度有所提高。使用该设备,我们还发现猫杯状病毒(FCV)的检出限为1.6 X 105 PFU / ml,这是人肠病毒的模型系统。该设备有潜力用作环境水样中此类肠道病毒的早期检测系统。

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    Kondapalli Sowmya;

  • 作者单位
  • 年度 2011
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  • 原文格式 PDF
  • 正文语种 en_US
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