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Lifetime of biomolecules in polymer-based hybrid nanodevices

机译：基于聚合物的混合纳米器件中生物分子的寿命

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Prolonging the lifetime of biomolecules in their functional states is critical for many applications where biomolecules are integrated into synthetic materials or devices. A simplified molecular shuttle system, which consists of fluorescently labelled microtubules propelled by kinesin motor proteins bound to the surface of a flowcell, served here as a model system to probe the lifetime of a hybrid device. In this system, the functional decay can easily be assayed by utilizing optical microscopy to detect motility and disintegration of microtubules. We found that the lifetimes of these hybrid systems were mainly limited by the stability of microtubules (MTs), rather than of kinesin. To determine the biocompatibility of polymers widely used in microfabrication, we assembled flowcells with glass bottom surfaces and covers fabricated from glass, poly(urethane) (PU), poly(methyl-methacrylate) (PMMA), poly (dimethyl siloxane) (PDMS) and ethylene-vinyl alcohol copolymer (EVOH). Without illumination, only PU had a substantial negative impact on MT stability, while PMMA, PDMS and EVOH showed stabilities comparable to glass. Under the influence of light, however, the MTs degraded rapidly in the presence of PDMS or PMMA, even in the presence of oxygen scavengers. A similar effect was observed on glass if oxygen scavengers were not added to the medium. Strong bleaching of the fluorophores was again only found on the polymer substrates and photobleaching coincided with an accelerated depolymerization of the MTs.
机译：对于许多将生物分子整合到合成材料或设备中的应用，延长其功能状态下的生物分子的寿命至关重要。一种简化的分子穿梭系统，由结合到流通池表面的驱动蛋白运动蛋白推动的荧光标记微管组成，在这里用作模型系统，以探测混合设备的寿命。在该系统中，可以通过利用光学显微镜检测微管的运动性和崩解性来轻松地检测功能衰退。我们发现这些混合系统的寿命主要受微管（MTs）而不是驱动蛋白的稳定性的限制。为了确定在微细加工中广泛使用的聚合物的生物相容性，我们组装了带有玻璃底面和盖的流通池，该底池由玻璃，聚氨脂（PU），聚甲基丙烯酸甲酯（PMMA），聚二甲基硅氧烷（PDMS）制成和乙烯-乙烯醇共聚物（EVOH）。没有照明，只有PU对MT稳定性有很大的负面影响，而PMMA，PDMS和EVOH的稳定性与玻璃相当。但是，在光的影响下，即使在存在PDMS或PMMA的情况下，甚至在存在除氧剂的情况下，MT也会迅速降解。如果不向介质中添加除氧剂，则会在玻璃上观察到类似的效果。再次仅在聚合物基材上发现了荧光团的强烈漂白，并且光漂白与MT的加速解聚同时发生。

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