To mimic organelles and cells and to construct next-generation therapeutics, asymmetric functionalization and location of proteins for artificial vesicles is thoroughly needed to emphasize the complex interplay of biological units and systems through spatially separated and spatiotemporal controlled actions, release, and communications. For the challenge of vesicle (= polymersome) construction, the membrane permeability and the location of the cargo are important key characteristics that determine their potential applications. Herein, an in situ and post loading process of avidin in pH-responsive and photo-cross-linked polymersomes is developed and characterized. First, loading efficiency, main location (inside, lumen, outside), and release of avidin under different conditions have been validated, including the pH-stable presence of avidin in polymersomes’ membrane outside and inside. This advantageous approach allows us to selectively functionalize the outer and inner membranes as well as the lumen with several bio(macro)molecules, generally suited for the construction of asymmetrically functionalized artificial organelles. In addition, a fluorescence resonance energy transfer (FRET) effect was used to study the permeability or uptake of the polymersome membrane against a broad range of biotinylated (macro)molecules (different typology, sizes, and shapes) under different conditions.

中文翻译：

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pH响应聚合物中的抗生物素蛋白定位，用于探测生物素化的（大分子）分子在膜和管腔中的对接。

为了模拟细胞器和细胞并构建下一代治疗剂，完全需要用于人工囊泡的蛋白质的不对称功能化和定位，以强调通过空间分隔和时空控制的动作，释放和通讯来实现生物单位和系统的复杂相互作用。对于囊泡（=聚合体）构造的挑战，膜的渗透性和货物的位置是决定其潜在应用的重要关键特征。在此，就地和后期开发并表征了亲和素在pH响应和光交联的聚合物中的负载过程。首先，已经验证了负载效率，主要位置（内部，内腔，外部）和抗生物素蛋白在不同条件下的释放，包括在聚合物囊体膜的内部和外部都存在pH稳定的抗生物素蛋白。这种有利的方法允许我们用几种生物（大分子）分子选择性地功能化外膜和内膜以及管腔，这些分子通常适用于不对称功能化的人造细胞器的构建。另外，使用荧光共振能量转移（FRET）效应研究了在不同条件下聚合物囊膜对各种生物素化（大分子）分子（不同的类型，大小和形状）的渗透性或摄取。