Immobilization of biotinylated bacteriophages on biosensor surfaces
Immobilization of biotinylated bacteriophages on biosensor surfaces
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DOI：
10.1016/j.snb.2007.03.007
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发表时间：
2007-08-08
影响因子：
8.4
通讯作者：
Evoy, S.
中科院分区：
文献类型：
--
作者：
Gervals, L.;Gel, M.;Evoy, S.
噬菌体是一种识别细菌表面特定受体的病毒，它们与之结合并注入遗传物质。这种识别的特异性为生物传感器的发展开辟了非凡的可能性。报道了T4噬菌体在金表面的化学附着。这种附着利用了噬菌体衣壳头部的遗传生物素化，以及生物素/链亲和素系统的天然亲和力。首先描述了一种链霉亲和素固定化化学的发展，该化学可以最大限度地减少目标细菌的非特异性结合。然后报道了基因生物素化的T4噬菌体附着在这些链霉亲和素涂覆的表面上。与野生型噬菌体在裸金上的简单物理吸附相比，这种化学固定可使附着提高15倍。然后使用附着程序来研究生物素化噬菌体终止表面对宿主细菌生长的影响。这项评估是在一个电池-衬底阻抗传感装置中进行的。链霉亲和素介导的生物素化噬菌体的附着可显著延迟宿主细菌的生长长达17.2小时。相比之下，发现野生型噬菌体与链亲和素表面的非特异性结合导致的生长延迟较小，为13小时。(c) 2007 Elsevier B.V.。
Bacteriophages are viruses that recognize specific receptors on the bacterium surface to which they bind and inject genetic material. The specificity of this recognition opens remarkable possibilities for biosensor development. The chemical attachment of T4 bacteriophages onto,gold surfaces is being reported. This attachment leverages the genetic biotinylation of the capsid heads of bacteriophages, and the natural affinity of the biotin/streptavidin system. The development of a streptavidin-immobilization chemistry that minimizes non-specific binding of the target bacterium is first described. The attachment of genetically biotinylated T4 bacteriophages onto these streptavidin-coated surfaces is then reported. Such chemical immobilization results in a 15-fold improvement of attachment when compared to the simple physisorption of the wild-type phage onto bare gold. The attachment procedure was then used to investigate the effect of a biotinylated phage-terminated surface on the growth of the host bacteria. This assessment was conducted in an electric cell-substrate impedance sensing device. The streptavidin-mediated attachment of biotinylated phages significantly delays the growth of the host bacteria by up to 17.2h. In comparison, non-specific binding of wild-type phages onto the streptavidin surface is found to cause a lesser growth delay of 13 h. (c) 2007 Elsevier B.V. All rights reserved.