Lifetime of biomolecules in polymer-based hybrid nanodevices
Lifetime of biomolecules in polymer-based hybrid nanodevices
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DOI：
10.1088/0957-4484/15/10/008
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发表时间：
2004-10-01
期刊：
影响因子：
3.5
通讯作者：
Vogel, V
中科院分区：
文献类型：
--
作者：
Brunner, C;Ernst, KH;Vogel, V
延长其功能状态生物分子的寿命对于许多将生物分子整合到合成材料或设备中的应用至关重要。一个简化的分子班车系统，由荧光标记的微管组成，这些微管由动机运动蛋白与流循环表面结合在一起，在这里用作模型系统，用于探测混合设备的寿命。在该系统中，可以通过使用光学显微镜检测微管的运动性和分解来轻松测定功能衰减。我们发现，这些混合系统的寿命主要受微管（MT）而不是动力蛋白的稳定性限制。为了确定广泛用于微生物的聚合物的生物相容性，我们用玻璃底部表面和玻璃，聚氨酯（氨基甲酸酯）（PU），聚（甲基 - 甲基丙烯酸酯）（PMMA）（PMMA），聚（二甲基硅氧烷）（PDMS）（PDMS）（PDMS）（PDMS）（PDMS）（PDMS）（PDMS）（PDMS）组装了玻璃底部表面和盖子组装。和乙烯 - 乙烯醇共聚物（EVOH）。没有照明，只有PU对MT稳定性产生了重大负面影响，而PMMA，PDMS和EVOH的稳定性与玻璃相当。然而，在光的影响下，即使在存在氧气清除剂的情况下，MTS在PDMS或PMMA的存在下也迅速降解。如果未在培养基中添加氧气清除剂，则观察到玻璃的类似效果。仅在聚合物底物上发现荧光团的强烈漂白，并在光漂白上与MTS的加速去聚合相吻合。
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.