Polymer particles with antibody-like affinity, i.e., molecularly imprinted polymers, offer an ideal platform for biopharmaceutical virus purification. In recent years, attempts combining molecular imprinting technology with a variety of visualization and detection techniques have been reported for directly confirming the localized presence of the template. Direct target visualization is crucial for the characterization of molecularly imprinted polymers, especially if biological templates such as viruses are used. In the present study, for the first time the viral binding behavior at virus-imprinted polymers (VIPs) via stimulated emission depletion (STED) microscopy is shown by imaging individual, fluorescently labeled virus particles. STED microscopy achieves among various other super-resolution techniques the best temporal resolution at high spatial resolution. An innovative virus purification material selective for human adenovirus type 5 (AdV5) offered highly purified virus for the subsequent fluorescent labeling procedure, thus enabling STED imaging. Excellent binding affinities (150-fold higher versus control particles) and high selectivity toward the target virus (AdV5) were observed at those VIPs, even in competitive binding experiments with minute virus of mice using dual-label STED microscopy.

中文翻译：

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使用超高分辨率光学显微镜揭示混合核壳结构基质上的直接病毒结合。

具有抗体样亲和力的聚合物颗粒，即分子印迹聚合物，为生物药物病毒纯化提供了理想的平台。近年来，已经报道了将分子印迹技术与多种可视化和检测技术相结合的尝试，以直接确认模板的局部存在。直接目标可视化对于表征分子印迹聚合物至关重要，尤其是在使用生物模板（例如病毒）的情况下。在本研究中，通过对单个荧光标记的病毒颗粒进行成像，首次显示了通过受激发射耗竭（STED）显微镜在病毒印迹聚合物（VIP）上的病毒结合行为。STED显微镜在各种其他超分辨率技术中实现了高空间分辨率下的最佳时间分辨率。一种对人类5型腺病毒（AdV5）有选择性的创新病毒纯化材料为后续的荧光标记程序提供了高度纯化的病毒，从而实现了STED成像。在那些VIP上也观察到了极好的结合亲和力（相对于对照颗粒高150倍）和对目标病毒（AdV5）的高选择性，甚至在使用双标记STED显微镜对小鼠微小病毒进行竞争性结合实验中也是如此。