Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Key Laboratory of Molecular Biophysics of MOE, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Department of Chemical and Biomolecular Engineering, Department of Chemistry and Biochemistry, University of California, Los Angeles, CA, USA.
Methods Mol Biol. 2024;2793:131-141. doi: 10.1007/978-1-0716-3798-2_9.
Phage-nanomaterial conjugates are functional bio-nanofibers with various applications. While phage display can select for phages with desired genetically encoded functions and properties, nanomaterials can endow the phages with additional features at nanoscale dimensions. Therefore, combining phages with nanotechnology can construct bioconjugates with unique characteristics. One strategy for filamentous phages is to adsorb nanoparticles onto the side wall, composed of pVIII subunits, through electrostatic interactions. However, a noncovalent approach may cause offloading if the environment changes, potentially causing side effects especially for in vivo applications. Therefore, building stable phage-bioconjugates is an important need. We previously reported the construction of chimeric M13 phage conjugated with gold nanorods, named "phanorods," without weakening the binding affinity to the bacterial host cells. Herein, we give a detailed protocol for preparing the chimeric M13 phage and covalently conjugating gold nanorods to the phage.
噬菌体-纳米材料缀合物是具有多种应用的功能性生物纳米纤维。噬菌体展示技术可以选择具有所需遗传编码功能和特性的噬菌体,而纳米材料可以在纳米尺度上赋予噬菌体额外的特性。因此,将噬菌体与纳米技术结合可以构建具有独特特性的生物缀合物。丝状噬菌体的一种策略是通过静电相互作用将纳米颗粒吸附到由 pVIII 亚基组成的侧壁上。然而,如果环境发生变化,非共价方法可能会导致纳米颗粒脱落,这可能会产生副作用,特别是在体内应用中。因此,构建稳定的噬菌体-生物缀合物是一个重要需求。我们之前报道了一种金纳米棒偶联的嵌合 M13 噬菌体的构建,命名为“phanorods”,其与细菌宿主细胞的结合亲和力没有减弱。本文详细介绍了制备嵌合 M13 噬菌体和将金纳米棒共价偶联到噬菌体上的方法。