An integrated self-assembled nanofluidic system for controlled biological chemistries
An integrated self-assembled nanofluidic system for controlled biological chemistries
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DOI：
10.1002/anie.200801606
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发表时间：
2008-01-01
影响因子：
16.6
通讯作者：
Vogel, Horst
中科院分区：
文献类型：
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作者：
Bolinger, Pierre-Yves;Stamou, Dimitrios;Vogel, Horst
强大的生物技术驱动的小型化反应系统，以减少样品消耗和增加吞吐量，到目前为止，主要是解决了微制造。[1-4]在本文中，我们使用自组装（SA）[5，6]来创建用于在封闭的毫微微升反应器容器（较大的单层脂质囊泡）中混合阿升体积（从纳米尺寸的脂质囊泡释放）的纳米流体系统，从而以单分子精度控制混合反应物的数量。通过荧光相关光谱（FCS）原位监测反应。反应物的混合，这引发了酶催化的非荧光底物转化为荧光产物，是由温度的变化，驱动小泡通过相变触发。[7，8]封闭的自主纳米反应器允许重复添加反应物和连续的不同反应;系统保持紧密密封数周。这种方法为化学和蛋白质文库的超小型化筛选、合成化学和生物学以及人工细胞的构建开辟了新的前景。生物细胞内的化学反应发生在从几微米到纳米的3D长度尺度上，这取决于细胞及其细胞内隔室的尺寸。生物系统采用分子SA作为一种普遍存在的方法来创建高度复杂和功能有效的纳米级架构，用于建立细胞生物化学网络。[5a]生物的微型化和生物化学过程的复杂性在实验室合成化学中是无与伦比的。例如，引发化学反应的最重要的实验室操作是在限定尺寸的反应器内混合限定体积和浓度的反应物。如果混合应该缩小到亚飞升体积，那么在宏观尺度上微不足道的事情就变成了一个未解决的挑战。目前的微技术能够处理小至50 fL的体积。[1-4]或者，由多肽、聚合物或脂质制成的自组装囊泡已经进化成包围
The strong biotechnological drive for miniaturized reaction systems to reduce sample consumption and increase throughput has, to date, been mainly addressed by microfabrication.[1–4] Herein, we use self-assembly (SA)[5, 6] to create a nanofluidic system for mixing attoliter volumes (released from nanometer-sized lipid vesicles) in a closed femtoliter reactor vessel (a larger unilamellar lipid vesicle), thereby controlling the number of mixed reactants with singlemolecule precision. The reactions are monitored in situ by fluorescence correlation spectroscopy (FCS). The mixing of reactants, which initiates an enzyme-catalyzed transformation of nonfluorescent substrate to a fluorescent product, is triggered by changes in temperature that drive the small vesicles through phase transitions.[7, 8] The closed, autonomous nanoreactor allows repetitive addition of reactants and successive distinct reactions; the system remains tightly sealed for weeks. This approach opens novel vistas for ultraminiaturized screening of chemical and protein libraries, synthetic chemistry and biology, and for constructing artificial cells.Chemical reactions inside biological cells occur on 3D length scales ranging from several micrometers to nanometers, as dictated by the dimensions of cells and their intracelluar compartments. Biological systems employ molecular SA as a ubiquitous method to create highly complex and functionally efficient nanoscale architectures for establishing the cellular biochemical network.[5a] The biological miniaturization and complexity of biochemical processes is unmatched in laboratory synthetic chemistry. For example, the most important laboratory manipulation to initiate a chemical reaction is the mixing of defined volumes and concentrations of reactants inside a reactor of defined size. What is trivial at macroscopic scale becomes an unsolved challenge if mixing should be downscaled to sub-femtoliter volumes. Present microtechnologies are capable of handling volumes as small as 50 fL.[1–4] Alternatively, self-assembled vesicles made of polypeptides, polymers, or lipids have evolved to enclose