Department of Biomedical Engineering, Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands; Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands.
Department of Biomedical Engineering, Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands; Department of Chemical Engineering and Chemistry, Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven, The Netherlands.
Methods Enzymol. 2021;646:51-82. doi: 10.1016/bs.mie.2020.06.008. Epub 2020 Jul 9.
The utilization of liquid-liquid phase separated systems has seen increased attention as synthetic cell platforms due to their innate ability to sequester interesting, functional, and biologically relevant materials. However, their applications are limited by the temporal stability of such condensed phases. While there are a number of strategies toward droplet stabilization, in our group we have developed a polymer-based approach to stabilize complex coacervate microdroplets. These protocells are remarkably robust and have been utilized to support a number of new protocellular applications. Here, we describe in detail the methodologies we have developed for the synthesis of the starting components, their formation into stable, cargo-loaded protocells, and how these protocells are treated post-formation to purify and analyze the resultant functional self-assembled systems.
由于液-液相分离系统具有内在的隔离有趣、功能和生物相关材料的能力，因此作为合成细胞平台，它们受到了越来越多的关注。然而，它们的应用受到这些凝聚相的时间稳定性的限制。虽然有许多策略可以稳定液滴，但我们小组开发了一种基于聚合物的方法来稳定复杂凝聚相微液滴。这些原细胞非常坚固，并已被用于支持许多新的原细胞应用。在这里，我们详细描述了我们开发的用于合成起始成分的方法、将其形成稳定的、载有货物的原细胞的方法，以及如何在形成后处理这些原细胞以纯化和分析所得功能自组装系统的方法。
