Koch Claudia, Poghossian Arshak, Schöning Michael J, Wege Christina
Institute of Biomaterials and Biomolecular Systems, University of Stuttgart, 70569 Stuttgart, Germany.
Institute of Nano- and Biotechnologies, FH Aachen, Campus Jülich, 52428 Jülich, Germany.
Nanotheranostics. 2018 Feb 20;2(2):184-196. doi: 10.7150/ntno.22114. eCollection 2018.
The presentation of enzymes on viral scaffolds has beneficial effects such as an increased enzyme loading and a prolonged reusability in comparison to conventional immobilization platforms. Here, we used modified tobacco mosaic virus (TMV) nanorods as enzyme carriers in penicillin G detection for the first time. Penicillinase enzymes were conjugated with streptavidin and coupled to TMV rods by use of a bifunctional biotin-linker. Penicillinase-decorated TMV particles were characterized extensively in halochromic dye-based biosensing. Acidometric analyte detection was performed with bromcresol purple as pH indicator and spectrophotometry. The TMV-assisted sensors exhibited increased enzyme loading and strongly improved reusability, and higher analysis rates compared to layouts without viral adapters. They extended the half-life of the sensors from 4 - 6 days to 5 weeks and thus allowed an at least 8-fold longer use of the sensors. Using a commercial budget-priced penicillinase preparation, a detection limit of 100 µM penicillin was obtained. Initial experiments also indicate that the system may be transferred to label-free detection layouts.
与传统固定化平台相比,在病毒支架上展示酶具有诸多有益效果,如增加酶负载量和延长可重复使用性。在此,我们首次将修饰后的烟草花叶病毒(TMV)纳米棒用作青霉素G检测中的酶载体。青霉素酶与链霉亲和素偶联,并通过双功能生物素连接子与TMV棒相连。在基于酸碱指示剂染料的生物传感中,对青霉素酶修饰的TMV颗粒进行了广泛表征。以溴甲酚紫作为pH指示剂,采用分光光度法进行酸碱分析物检测。与没有病毒适配体的布局相比,TMV辅助传感器表现出更高的酶负载量、显著提高的可重复使用性以及更高的分析速率。它们将传感器的半衰期从4 - 6天延长至5周,从而使传感器的使用时间至少延长了8倍。使用商业预算价格的青霉素酶制剂,获得了100 μM青霉素的检测限。初步实验还表明,该系统可转换为无标记检测布局。