Karig David K, Bessling Seneca, Thielen Peter, Zhang Sherry, Wolfe Joshua
Research and Exploratory Development Department, Johns Hopkins University Applied Physics Laboratory, Laurel, MD, USA
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA.
J R Soc Interface. 2017 Apr;14(129). doi: 10.1098/rsif.2016.1039.
Many biotechnology capabilities are limited by stringent storage needs of reagents, largely prohibiting use outside of specialized laboratories. Focusing on a large class of protein-based biotechnology applications, we address this issue by developing a method for preserving cell-free protein expression systems for months above room temperature. Our approach realizes unprecedented long-term stability at elevated temperatures by leveraging the sugar alcohol trehalose, a simple, low-cost, open-air drying step, and strategic separation of reaction components during drying. The resulting preservation capacity enables efficient production of a wide range of on-demand proteins under adverse conditions, for instance during emergency outbreaks or in remote locations. To demonstrate application potential, we use cell-free reagents subjected to months of exposure at 37°C and atmospheric conditions to produce sufficient concentrations of a pyocin protein to kill , a troublesome pathogen for traumatic and burn wound injuries. Our work makes possible new biotechnology applications that demand ruggedness and scalability.
许多生物技术能力受到试剂严格储存需求的限制,这在很大程度上禁止了其在专业实验室之外的使用。针对一大类基于蛋白质的生物技术应用,我们通过开发一种在室温以上保存无细胞蛋白质表达系统数月的方法来解决这一问题。我们的方法通过利用糖醇海藻糖、简单的低成本露天干燥步骤以及干燥过程中反应成分的策略性分离,在高温下实现了前所未有的长期稳定性。由此产生的保存能力能够在不利条件下高效生产各种按需蛋白质,例如在紧急疫情爆发期间或偏远地区。为了证明其应用潜力,我们使用在37°C和大气条件下暴露数月的无细胞试剂来生产足够浓度的绿脓菌素蛋白,以杀死铜绿假单胞菌,这是一种对创伤和烧伤伤口造成麻烦的病原体。我们的工作使那些需要坚固性和可扩展性的新生物技术应用成为可能。