Highly Active Protein Surfaces Enabled by Plant-Based Polyphenol Coatings.
Highly Active Protein Surfaces Enabled by Plant-Based Polyphenol Coatings.
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DOI：
10.1021/acsami.8b13793
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发表时间：
2018-10
影响因子：
9.5
通讯作者：
Ana M. L. Sousa;Tai-De Li;Sabu Varghese;P. Halling;King Hang Aaron Lau
中科院分区：
文献类型：
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作者：
Ana M. L. Sousa;Tai-De Li;Sabu Varghese;P. Halling;King Hang Aaron Lau
蛋白质代表复杂的生物分子，能够广泛但也具有高度特异性的功能。它们在材料载体上的固定化可以实现从传感和工业生物催化到生物医学界面和材料的广泛应用。我们证明了使用水性处理的交联多酚涂层用于将蛋白质（包括IgG、抗生物素蛋白和各种单结构域和多结构域酶）固定在不同材料上以实现活性生物功能结构（例如，约在纳米多孔纤维素和氧化铝、钢丝网和聚酯织物上分别为2.2、1.7、1.1和4.8 mg·m-2活性磷酸酶）。酶测定，X-射线光电子能谱，银染色，补充接触角，固态13 C NMR，HPLC和ESI-MS测量被用来表征多酚，涂层，和蛋白质层。我们表明，功能化过程可以有利地直接针对蛋白质活性进行优化，而不是传统地关注涂层的厚度。多酚涂层比其他方法（如聚多巴胺）具有更高的活性（在某些情况下超过一个数量级）和更宽的工艺pH值和材料相容性。由不同的植物多酚提取物形成的涂层，即使在较低的纯度（和成本）下，也被发现是高度功能性的。化学上，我们的研究结果表明，多酚涂层不同于聚多巴胺，主要是因为消除了胺基，多酚层与中间水平的反应性可能会更好地导致高固定化蛋白质的活性。总的来说，已经获得了对简单易用的多酚涂层的更好的理解，这使得可以应用于不同材料和纳米结构支持物的活性蛋白质表面的显着发展成为可能。
Proteins represent complex biomolecules capable of wide-ranging but also highly specific functionalities. Their immobilization on material supports can enable broad applications from sensing and industrial biocatalysis to biomedical interfaces and materials. We demonstrate the advantages of using aqueous-processed cross-linked polyphenol coatings for immobilizing proteins, including IgG, avidin, and various single and multidomain enzymes on diverse materials, to enable active biofunctional structures (e.g., ca. 2.2, 1.7, 1.1, and 4.8 mg·m-2 active phosphatase on nanoporous cellulose and alumina, steel mesh, and polyester fabric, respectively). Enzyme assays, X-ray photoelectron spectroscopy, silver staining, supplemented with contact angle, solid-state 13C NMR, HPLC, and ESI-MS measurements were used to characterize the polyphenols, coatings, and protein layers. We show that the functionalization process may be advantageously optimized directly for protein activity rather than the traditional focus on the thickness of the coating layer. Higher activities (by more than an order of magnitude in some cases) and wider process pH and material compatibility are demonstrated with polyphenol coatings than other approaches such as polydopamine. Coatings formed from different plant polyphenol extracts, even at lowered purity (and cost), were also found to be highly functional. Chemically, our results indicate that polyphenol coatings differ from polydopamine mainly because of the elimination of amine groups, and that polyphenol layers with intermediate levels of reactivity may better lead to high immobilized protein activity. Overall, an improved understanding of simple-to-use polyphenol coatings has been obtained, which enabled a significant development in active protein surfaces that may be applied across diverse materials and nanostructured supports.