Production of tunable nanomaterials using hierarchically assembled bacteriophages
Production of tunable nanomaterials using hierarchically assembled bacteriophages
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DOI：
10.1038/nprot.2017.085
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发表时间：
2017-09-01
期刊：
影响因子：
14.8
通讯作者：
Lee, Seung-Wuk
中科院分区：
文献类型：
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作者：
Lee, Ju Hun;Warner, Christopher M.;Lee, Seung-Wuk
大规模制造具有可调功能的精确定义的纳米结构对于利用纳米科学和纳米技术生产电子器件、能量发生器、生物传感器和生物医学至关重要。虽然自组装技术已经被开发出来用于开发生物分子作为功能材料，但由此产生的纳米结构和功能仍然非常有限，而且还远未实现可扩展的合成。最近，我们建立了一种以噬菌体为基础的仿生过程，称为“自我模板组装”。我们使用噬菌体作为纳米纤维模型系统来利用其液晶结构来创建不同的层级组织结构。我们还证明了噬菌体功能肽的遗传修饰可以产生可用作软组织和硬组织再生材料、生物传感器和能源材料的结构。在这里，我们描述了一种进行噬菌体基因工程、滴度放大、纯化和自模板组装的综合方案，并提出了使用光学显微镜、原子力显微镜(AFM)和扫描电子显微镜(SESEM)表征分级噬菌体纳米结构的方法。我们还讨论了污染源，制造过程中的常见错误，以及确保可再生材料生产的质量控制措施。该方案大约需要8-10天才能完成。
Large-scale fabrication of precisely defined nanostructures with tunable functions is critical to the exploitation of nanoscience and nanotechnology for production of electronic devices, energy generators, biosensors, and bionanomedicines. Although self-assembly processes have been developed to exploit biological molecules for functional materials, the resulting nanostructures and functions are still very limited, and scalable synthesis is far from being realized. Recently, we have established a bacteriophage-based biomimetic process, called 'self-templating assembly'. We used bacteriophage as a nanofiber model system to exploit its liquid crystalline structure for the creation of diverse hierarchically organized structures. We have also demonstrated that genetic modification of functional peptides of bacteriophage results in structures that can be used as soft and hard tissue-regenerating materials, biosensors, and energy-generating materials. Here, we describe a comprehensive protocol to perform genetic engineering of phage, liter-scale amplification, purification, and self-templating assembly, and suggest approaches for characterizing hierarchical phage nanostructures using optical microscopy, atomic-force microscopy (AFM), and scanning electron microscopy (SESEM). We also discuss sources of contamination, common mistakes during the fabrication process, and quality-control measures to ensure reproducible material production. The protocol takes similar to 8-10 d to complete.