3d localization and diffusion of proteins in polyelectrolyte multilayers
3d localization and diffusion of proteins in polyelectrolyte multilayers
复制标题
聚电解质多层中蛋白质的 3D 定位和扩散
作者：
Madaboosi;Schmidt;Duschl;Volodkin
各种生物材料与表面的相互作用比以往任何时候都更加重要，以确保生物医学应用的效率和可重复性。非常有趣的表面材料是微米厚的聚电解质多层材料。不仅它们的表面，而且它们的主体也可以装载生物材料，如蛋白质或DNA，用于各种目的。因此，我们建立了一种用激光共聚焦扫描显微镜分析由聚赖氨酸和透明质酸组成的聚电解质膜中不同大小和电荷的荧光标记蛋白的横向和纵向分布的方法。这种方法使我们能够通过光漂白后的荧光恢复来测量蛋白质的扩散系数，作为它们在膜中的垂直位置的函数，并有助于在空间和时间上以高分辨率理解膜中的分子相互作用。因此，我们证实了膜中的蛋白质负载是由静电相互作用驱动的——10和500 kDa的不带电葡聚糖分子不会扩散到膜中。不同尺寸的蛋白质（3 ~ 11 nm）与净电荷无关，其扩散速度相对较快（D = 2 ~ 4 μm2 s−1），表明聚合物间相互作用复杂。该方法通过发现分子间相互作用与生物分子对生物样品（如细胞）或检测单元（如生物传感器）的迁移性和可用性之间的关系，为设计用于生物应用的聚电解质多层膜提供了一种新的强大的实验工具。
The interaction of diverse biomaterials with surfaces is more crucial than ever for biomedical applications to ensure efficiency and reproducibility. Very interesting surface materials are micrometer-thick polyelectrolyte multilayers. Not only their surface but also the bulk can be loaded with biomaterials like proteins or DNA for various purposes. Therefore, we established a method to analyze the lateral and vertical distribution of fluorescently labelled proteins of various size and charge in polyelectrolyte films composed of poly(L-lysine) and hyaluronic acid by confocal laser scanning microscopy. This approach enables us to measure the diffusion coefficients of the proteins via fluorescence recovery after photobleaching as a function of their vertical position in the film and facilitates the understanding of molecular interactions in the film with a high resolution in both space and time. As a result, we confirm that protein loading in the film is driven by electrostatic interactions – uncharged dextran molecules of 10 and 500 kDa do not diffuse into the film. Proteins of different sizes (3–11 nm) can diffuse relatively fast (D = 2–4 μm2 s−1) independent of their net charge, indicating complex interpolymer interactions. This approach is a new powerful experimental tool to design the polyelectrolyte multilayers for bio-applications by finding a relationship between intermolecular interactions and mobility and availability of biomolecules to biological samples (e.g. cells) or detection units (e.g. biosensors).
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微流体技术与软层层薄膜的结合：3D 聚合物结构中的选择性细胞生长。
影响因子：
6.1
作者：
N. Madaboosi;K. Uhlig;Stephan Schmidt;M. Jäger;H. Möhwald;C. Duschl;D. Volodkin
通讯作者：
D. Volodkin
影响因子：
4.6
作者：
W. Ouyang;M. Müller
通讯作者：
W. Ouyang;M. Müller
影响因子：
3.4
作者：
Armstrong, JK;Wenby, RB;Fisher, TC
通讯作者：
Fisher, TC
影响因子：
3.9
作者：
Jourdainne, Laurent;Lecuyer, Sigolene;Charitat, Thierry
通讯作者：
Charitat, Thierry
影响因子：
13.3
作者：
Richard, Doriane;Nguyen, Isabelle;Ogier, Joelle
通讯作者：
Ogier, Joelle