Monomeric streptavidin phage display allows efficient immobilization of bacteriophages on magnetic particles for the capture, separation, and detection of bacteria.
Monomeric streptavidin phage display allows efficient immobilization of bacteriophages on magnetic particles for the capture, separation, and detection of bacteria.
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DOI：
10.1038/s41598-023-42626-9
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发表时间：
2023-09-27
影响因子：
4.6
通讯作者：
Nugen, Sam R.
中科院分区：
文献类型：
--
作者：
Carmody, Caitlin M.;Nugen, Sam R.
将噬菌体固定到固体支持物如磁性颗粒上已经证明了作为用于分离和检测其宿主细菌的生物传感器的超低检测限。虽然磁化细菌的潜在影响很大，但目前的固定方法要么是弱的、昂贵的、低效的，要么是费力的，使得它们不太适合商业化。为了弥合这一差距，我们开发了一种高效、位点特异性和低成本的方法来将噬菌体固定在固体载体上。虽然链霉亲和素-生物素代表了理想的缀合方法，但用链霉亲和素官能化磁性颗粒需要平方米的覆盖范围，因此不适合低成本测定。在这里，我们通过基因工程改造噬菌体，使其能够在感染细菌宿主的过程中合成单体链霉亲和素。单体链霉亲和素融合到衣壳蛋白（Hoc），以允许每个衣壳多达155个融合蛋白的位点特异性自组装。在E.大肠杆菌检测试验，检测限为100 mL水中< 10 CFU。这项工作突出了具有新型衣壳修饰的遗传修饰噬菌体的创造，扩大了噬菌体功能化生物技术的潜力。
Immobilization of bacteriophages onto solid supports such as magnetic particles has demonstrated ultralow detection limits as biosensors for the separation and detection of their host bacteria. While the potential impact of magnetized phages is high, the current methods of immobilization are either weak, costly, inefficient, or laborious making them less viable for commercialization. In order to bridge this gap, we have developed a highly efficient, site-specific, and low-cost method to immobilize bacteriophages onto solid supports. While streptavidin–biotin represents an ideal conjugation method, the functionalization of magnetic particles with streptavidin requires square meters of coverage and therefore is not amenable to a low-cost assay. Here, we genetically engineered bacteriophages to allow synthesis of a monomeric streptavidin during infection of the bacterial host. The monomeric streptavidin was fused to a capsid protein (Hoc) to allow site-specific self-assembly of up to 155 fusion proteins per capsid. Biotin coated magnetic nanoparticles were functionalized with mSA-Hoc T4 phage demonstrated in an E. coli detection assay with a limit of detection of < 10 CFU in 100 mLs of water. This work highlights the creation of genetically modified bacteriophages with a novel capsid modification, expanding the potential for bacteriophage functionalized biotechnologies.
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影响因子：
5.8
作者：
Hosseinidoust, Zeinab;Olsson, Adam U.;Tufenkji, Nathalie
通讯作者：
Tufenkji, Nathalie
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
DOI：
10.1073/pnas.0601211103
发表时间：
2006-03-28
影响因子：
11.1
作者：
Edgar, R;McKinstry, M;Adhya, S
通讯作者：
Adhya, S
DOI：
10.3390/s90806298
发表时间：
2009
期刊：
Sensors (Basel, Switzerland)
影响因子：
--
作者：
Archer MJ;Liu JL
通讯作者：
Liu JL
影响因子：
4.2
作者：
Arya, Sunil K.;Singh, Amit;Evoy, S.
通讯作者：
Evoy, S.