Center for Micro-Engineered Materials and the Department of Chemical and Biological Engineering, The University of New Mexico, Albuquerque, New Mexico 87131, United States.
Department of Internal Medicine, Molecular Medicine, The University of New Mexico, Albuquerque, New Mexico 87131, United States.
ACS Nano. 2022 Feb 22;16(2):2164-2175. doi: 10.1021/acsnano.1c08103. Epub 2022 Feb 10.
Preservation of evolved biological structure and function in robust engineering materials is of interest for storage of biological samples before diagnosis and development of vaccines, sensors, and enzymatic reactors and has the potential to avoid cryopreservation and its associated cold-chain issues. Here, we demonstrate that "freezing cells in amorphous silica" is a powerful technique for long-term preservation of whole mammalian cell proteomic structure and function at room temperature. Biomimetic silicification employs the crowded protein microenvironment of mammalian cells as a catalytic framework to proximally transform monomeric silicic acid into silicates forming a nanoscopic silica shell over all biomolecular interfaces. Silicification followed by dehydration preserves and passivates proteomic information within a nanoscale thin silica coating that exhibits size selective permeability (<3.6 nm), preventing protein leaching and protease degradation of cellular contents, while providing access of small molecular constituents for cellular enzymatic reaction. Exposure of dehydrated silicified cells to mild etchant or prolonged hydrolysis removes the silica, completely rerevealing biomolecular components and restoring their accessibility and functionality.
在进行诊断和疫苗、传感器、酶反应器开发之前,保存生物样本的进化生物结构和功能,这对存储生物学样本很重要,并且有避免冷冻保存及其相关冷链问题的潜力。在这里,我们证明了“将细胞冷冻在无定形二氧化硅中”是一种在室温下长期保存完整哺乳动物细胞蛋白质组结构和功能的强大技术。仿生硅化作用利用哺乳动物细胞拥挤的蛋白质微环境作为催化框架,将单体硅酸近就地转化为硅酸盐,在所有生物分子界面上形成纳米级的二氧化硅壳。硅化后进行脱水处理,可以在纳米级薄的二氧化硅涂层内保存和钝化蛋白质组信息,该涂层具有尺寸选择性渗透性(<3.6nm),可以防止蛋白质浸出和细胞内容物中蛋白酶的降解,同时为细胞酶反应提供小分子成分的通道。将脱水硅化的细胞暴露于温和的蚀刻剂或长时间的水解中,可以去除二氧化硅,完全重新揭示生物分子成分,并恢复其可及性和功能。