Gehlbach Emma M, Robinson Abbey O, Engelhart Aaron E, Adamala Katarzyna P
University of Minnesota Department of Genetics, Cell Biology and Development, Minneapolis, MN, USA.
bioRxiv. 2023 Oct 15:2023.10.15.562404. doi: 10.1101/2023.10.15.562404.
Small, spherical vesicles are a widely used chassis for the formation of model protocells and investigating the beginning of compartmentalized evolution. Various methods exist for their preparation, with one of the most common approaches being gentle hydration, where thin layers of lipids are hydrated with aqueous solutions and gently agitated to form vesicles. An important benefit to gentle hydration is that the method produces vesicles without introducing any organic contaminants, such as mineral oil, into the lipid bilayer. However, compared to other methods of liposome formation, gentle hydration is much less efficient at encapsulating aqueous cargo. Improving the encapsulation efficiency of gentle hydration would be of broad use for medicine, biotechnology, and protocell research. Here, we describe a method of sequentially hydrating lipid thin films to increase encapsulation efficiency. We demonstrate that sequential gentle hydration significantly improves encapsulation of water-soluble cargo compared to the traditional method, and that this improved efficiency is dependent on buffer composition. Similarly, we also demonstrate how this method can be used to increase concentrations of oleic acid, a fatty acid commonly used in origins of life research, to improve the formation of vesicles in aqueous buffer.
小型球形囊泡是一种广泛用于构建模型原始细胞和研究区室化进化起源的底盘。其制备方法多种多样，最常见的方法之一是温和水合，即脂质薄层与水溶液水合并轻轻搅拌以形成囊泡。温和水合的一个重要优点是该方法产生的囊泡不会将任何有机污染物（如矿物油）引入脂质双层。然而，与其他脂质体形成方法相比，温和水合在包封水性物质方面的效率要低得多。提高温和水合的包封效率在医学、生物技术和原始细胞研究中具有广泛的用途。在这里，我们描述了一种顺序水合脂质薄膜以提高包封效率的方法。我们证明，与传统方法相比，顺序温和水合显著提高了水溶性物质的包封率，并且这种提高的效率取决于缓冲液组成。同样，我们还展示了该方法如何用于提高油酸（生命起源研究中常用的一种脂肪酸）的浓度，以促进在水性缓冲液中形成囊泡。
