Shivakumar Tejasvi, Clark Joshua, Goode Alice, Anyanwu Valentine E, Williams Philip M
Molecular Therapeutics and Formulation, School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK.
Bioengineering (Basel). 2025 Feb 22;12(3):223. doi: 10.3390/bioengineering12030223.
Centralised cell-based biomanufacturing severely limits applicability in low-resource and extreme environments, where a largely untreated human population is present. Cell-free protein synthesis (CFPS) can surpass many of these limitations, due to its flexibility and low maintenance. After initial optimisation for high-level expression, we conceptualised CFPS platforms composed of lyophilised pellets and cellulose stacks for ease of storage and distribution. The latter platform consisted of lyophilised components on cellulose discs, which were layered and rehydrated to kickstart protein synthesis. Such paper-encompassed reactions were capable of robust expression, where the system can be modulated by simply changing the DNA layer. Using an initial screening design followed by a minimalistic design of experiments approach, we were able to improve the shelf life of lyophilised CFPS at room temperature from <1 week to 100% preservation at month 1. We anticipate that our strategy will enable quicker and more efficient stability optimisation for sustainable applications in all environments.
基于细胞的集中式生物制造严重限制了其在资源匮乏和极端环境中的适用性，而这些地区存在大量未得到充分治疗的人群。无细胞蛋白质合成（CFPS）由于其灵活性和低维护要求，可以克服许多此类限制。在对高水平表达进行初步优化后，我们构思了由冻干颗粒和纤维素堆栈组成的CFPS平台，以便于储存和分发。后一种平台由纤维素圆盘上的冻干成分组成，这些成分被分层并重新水化以启动蛋白质合成。这种包含纸张的反应能够实现强劲表达，其中系统可以通过简单地改变DNA层来进行调节。通过采用初步筛选设计，随后采用简约的实验设计方法，我们能够将冻干CFPS在室温下的保质期从不到1周提高到第1个月时100%保存。我们预计，我们的策略将能够更快、更有效地优化稳定性，以实现所有环境中的可持续应用。
