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表面功能化对噬菌体覆盖表面的影响及其对固定化噬菌体的磁弹性生物传感器后续性能的影响。

Effects of surface functionalization on the surface phage coverage and the subsequent performance of phage-immobilized magnetoelastic biosensors.

机构信息

Materials Engineering Program, Department of Mechanical Engineering, Auburn University, Auburn, AL 36849, USA.

出版信息

Biosens Bioelectron. 2011 Jan 15;26(5):2361-7. doi: 10.1016/j.bios.2010.10.012. Epub 2010 Oct 14.

DOI:10.1016/j.bios.2010.10.012
PMID:21084182
Abstract

One of the important applications for which phage-immobilized magnetoelastic (ME) biosensors are being developed is the wireless, on-site detection of pathogenic bacteria for food safety and bio-security. Until now, such biosensors have been constructed by immobilizing a landscape phage probe on gold-coated ME resonators via physical adsorption. Although the physical adsorption method is simple, the immobilization stability and surface coverage of phage probes on differently functionalized sensor surfaces need to be evaluated as a potential way to enhance the detection capabilities of the biosensors. As a model study, a filamentous fd-tet phage that specifically binds streptavidin was adsorbed on either bare or surface-functionalized gold-coated ME resonators. The surface functionalization was performed through the formation of three self-assembled monolayers with a different terminator, based on the sulfur-gold chemistry: AC (activated carboxy-terminated), ALD (aldehyde-terminated), and MT (methyl-terminated). The results, obtained by atomic force microscopy, showed that surface functionalization has a large effect on the surface phage coverage (46.8%, 49.4%, 4.2%, and 5.2% for bare, AC-, ALD-, and MT-functionalized resonators, respectively). In addition, a direct correlation of the observed surface phage coverage with the quantity of subsequently captured streptavidin-coated microbeads was found by scanning electron microscopy and by resonance frequency measurements of the biosensors. The differences in surface phage coverage on the differently functionalized surfaces may then be used to pattern the phage probe layer onto desired parts of the sensor surface to enhance the detection capabilities of ME biosensors.

摘要

噬菌体固定磁弹(ME)生物传感器的一个重要应用是无线、现场检测食品安全性和生物安全性的致病性细菌。到目前为止,这种生物传感器是通过物理吸附将景观噬菌体探针固定在金涂 ME 谐振器上来构建的。尽管物理吸附法简单,但需要评估不同功能化传感器表面上噬菌体探针的固定稳定性和表面覆盖率,这是增强生物传感器检测能力的一种潜在方法。作为模型研究,一种专门结合链霉亲和素的丝状 fd-tet 噬菌体通过形成三种不同终止剂的自组装单层来吸附在裸金涂 ME 谐振器或表面功能化的金涂 ME 谐振器上,基于硫金化学:AC(激活羧基末端)、ALD(醛基末端)和 MT(甲基末端)。原子力显微镜的结果表明,表面功能化对表面噬菌体覆盖率有很大影响(裸、AC-、ALD-和 MT-功能化谐振器的表面噬菌体覆盖率分别为 46.8%、49.4%、4.2%和 5.2%)。此外,通过扫描电子显微镜和生物传感器的共振频率测量发现,观察到的表面噬菌体覆盖率与随后捕获的链霉亲和素包被的微球的数量之间存在直接相关性。然后,可以利用不同功能化表面上的表面噬菌体覆盖率差异,将噬菌体探针层图案化到传感器表面的所需部分,以增强 ME 生物传感器的检测能力。

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