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Development of phage/antibody immobilized magnetostrictive biosensors.

机译：噬菌体/抗体固定磁致伸缩生物传感器的开发。

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摘要

There is an urgent need for biosensors that are able to detect and quantify the presence of a small amount of pathogens in a real-time manner accurately and quickly to guide prevention efforts and assay food and water quality. Acoustic wave (AW) devices, whose performance is defined by mass sensitivity (Sm) and quality factor (Q value), have been extensively studied as high performance biosensor platforms. However, current AW devices still face some challenges such as the difficulty to be employed in liquid and low Q value in practical applications. The objective of this research is to develop magnetostrictive sensors which include milli/microcantilever type (MSMC) and particle type (MSP).;Compared to other AW devices, MSMC exhibits the following advantages: (1) wireless/remote driving and sensing; (2) easy to fabricate; (3) works well in liquid; (4) exhibits a high Q value (> 500 in air). The fundamental study of the damping effect on MSMCs from the surrounding media including air and liquids were conducted to improve the Q value of MSMCs. The experiment results show that the Q value is dependent on the properties of surrounding media (e.g. viscosity, density), the geometry of the MSMCs, and the harmonic mode on the resonance behavior of MSMCs, etc.;The phage-coated MSMC has high specificity and sensitivity even while used in water with a low concentration of targeted bacteria. Two currently developed phages, JRB7 and E2, respectively respond to Bacillus anthracis spores and Salmonella typhimurium, were employed as bio-recognition elements in this research. The phage-immobilized MSMC biosensors exhibited high performance and detection of limit was 5 x 104 cfu/ml for the MSMC in size of 1.4 x 0.8 x 0.035 mm. The MSMC-based biosensors were indicated as a very potential method for in-situ monitoring of the biological quality in water.;The MSP combine antibody was used to detect Staphylococcus aureus in this experiment. The interface between MSPs and antibody was modified using Traut's Reagent by introducing the sulfhydryl group. To improve the mass sensitivity of magnetostrictive biosensors, several blocking agents were used to resist the nonspecific adsorption of S. aureus on the surface of the magnetostrictive biosensors and the blocking effects were studied by using ELISA and SEM. The results showed casein was one of the best blocking agents to resist the nonspecific binding in this experiment. Casein blocked antibody immobilized MSP biosensors exhibited high sensitivity and the limit of detection is 102 cfu/ml.
机译：迫切需要能够实时，准确，快速地检测和定量少量病原体的生物传感器，以指导预防工作并分析食品和水质。声波（AW）设备的性能由质量灵敏度（Sm）和品质因数（Q值）定义，已被作为高性能生物传感器平台进行了广泛的研究。然而，当前的AW器件仍然面临一些挑战，例如在液体应用中的难度以及在实际应用中的低Q值。这项研究的目的是开发包括毫微/微悬臂式（MSMC）和颗粒式（MSP）的磁致伸缩传感器。与其他AW设备相比，MSMC具有以下优点：（1）无线/远程驱动和传感； （2）易于制造； （3）在液体中效果良好； （4）具有很高的Q值（空气中> 500）。对包括空气和液体在内的周围介质对MSMC的阻尼作用进行了基础研究，以提高MSMC的Q值。实验结果表明，Q值取决于周围介质的性质（例如粘度，密度），MSMC的几何形状以及谐波模式对MSMCs共振行为的影响等。即使在目标细菌浓度低的水中使用时也具有特异性和敏感性。目前研究中使用的两种噬菌体JRB7和E2分别对炭疽芽孢杆菌孢子和鼠伤寒沙门氏菌有反应，被用作本研究的生物识别元件。固定有噬菌体的MSMC生物传感器表现出高性能，对于尺寸为1.4 x 0.8 x 0.035 mm的MSMC，极限检测为5 x 104 cfu / ml。基于MSMC的生物传感器被认为是一种非常有潜力的原位监测水中生物质量的方法。该实验中使用MSP组合抗体检测金黄色葡萄球菌。通过引入巯基，使用Traut试剂对MSP和抗体之间的界面进行了修饰。为了提高磁致伸缩生物传感器的质量灵敏度，使用了几种封闭剂来抵抗金黄色葡萄球菌在磁致伸缩生物传感器表面的非特异性吸附，并通过ELISA和SEM研究了其封闭效果。结果表明，在该实验中，酪蛋白是抵抗非特异性结合的最佳阻断剂之一。固定酪蛋白的抗体固定化MSP生物传感器表现出高灵敏度，检测限为102 cfu / ml。

著录项

  • 作者

    Fu, Liling.;

  • 作者单位

    Auburn University.;

  • 授予单位 Auburn University.;
  • 学科 Engineering Biomedical.;Engineering Mechanical.;Engineering Materials Science.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 209 p.
  • 总页数 209
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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