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基于噬菌体的光流控环形谐振器中无标记生物分子检测

Phage-based label-free biomolecule detection in an opto-fluidic ring resonator.

作者信息

Zhu Hongying, White Ian M, Suter Jonathan D, Fan Xudong

机构信息

Department of Biological Engineering, University of Missouri, 240D Bond Life Sciences Center, Columbia, MO 65211, United States.

出版信息

Biosens Bioelectron. 2008 Nov 15;24(3):461-6. doi: 10.1016/j.bios.2008.04.028. Epub 2008 May 9.

DOI:10.1016/j.bios.2008.04.028
PMID:18550355
Abstract

We have developed a sensitive and inexpensive opto-fluidic ring resonator (OFRR) biosensor using phage as a receptor for analyte detection. Phages have distinct advantages over antibodies as biosensor receptors. First, affinity selection from large libraries of random peptides displayed on phage provides a generic method of discovering receptors for detecting a wide range of analytes with high specificity and sensitivity. Second, phage production can be less complicated and less expensive than antibody production. Third, phages withstand harsh environments, reducing the environmental limitations and enabling regeneration of the biosensor surface. In this work, filamentous phage R5C2, displaying peptides that bind streptavidin specifically, was employed as a model receptor to demonstrate the feasibility of a phage-based OFRR biosensor. The experimental detection limit was approximately 100pM streptavidin and the K(d(apparent)) is 25pM. Specificity was verified using the RAP 5 phage, which is not specific to streptavidin, as the negative control. Sensing surface regeneration results show that the phage maintained functionality after surface regeneration, which greatly improves the sensors' reusability. The phage-based OFRR biosensor will become a promising platform for universal biomolecule detection with high sensitivity, low cost, and good reusability.

摘要

我们开发了一种灵敏且廉价的光流体环形谐振器(OFRR)生物传感器,该传感器使用噬菌体作为受体来检测分析物。与抗体作为生物传感器受体相比,噬菌体具有明显的优势。首先,从展示在噬菌体上的大量随机肽库中进行亲和力选择,提供了一种通用方法,可用于发现能够以高特异性和灵敏度检测多种分析物的受体。其次,噬菌体的生产可能比抗体生产更简单且成本更低。第三,噬菌体能够耐受恶劣环境,减少了环境限制,并使生物传感器表面能够再生。在这项工作中,丝状噬菌体R5C2展示了能特异性结合链霉亲和素的肽,被用作模型受体来证明基于噬菌体的OFRR生物传感器的可行性。实验检测限约为100pM链霉亲和素,表观解离常数(K(d(apparent)))为25pM。使用对链霉亲和素无特异性的RAP 5噬菌体作为阴性对照来验证特异性。传感表面再生结果表明,噬菌体在表面再生后仍保持功能,这大大提高了传感器的可重复使用性。基于噬菌体的OFRR生物传感器将成为一个有前景的平台,用于高灵敏度、低成本且具有良好可重复使用性的通用生物分子检测。

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