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Characterization of lipid composition and diffusivity in OLA generated vesicles.
Biochimica et Biophysica Acta (BBA) - Biomembranes ( IF 2.8 ) Pub Date : 2020-05-13 , DOI: 10.1016/j.bbamem.2020.183359
Michael Schaich 1 , Diana Sobota 1 , Hannah Sleath 1 , Jehangir Cama 2 , Ulrich F Keyser 1
Affiliation  

Giant Unilamellar Vesicles (GUVs) are a versatile tool in many branches of science, including biophysics and synthetic biology. Octanol-Assisted Liposome Assembly (OLA), a recently developed microfluidic technique enables the production and testing of GUVs within a single device under highly controlled experimental conditions. It is therefore gaining significant interest as a platform for use in drug discovery, the production of artificial cells and more generally for controlled studies of the properties of lipid membranes. In this work, we expand the capabilities of the OLA technique by forming GUVs of tunable binary lipid mixtures of DOPC, DOPG and DOPE. Using fluorescence recovery after photobleaching we investigated the lateral diffusion coefficients of lipids in OLA liposomes and found the expected values in the range of 1 μm2/s for the lipid systems tested. We studied the OLA derived GUVs under a range of conditions and compared the results with electroformed vesicles. Overall, we found the lateral diffusion coefficients of lipids in vesicles obtained with OLA to be quantitatively similar to those in vesicles obtained via traditional electroformation. Our results provide a quantitative biophysical validation of the quality of OLA derived GUVs, which will facilitate the wider use of this versatile platform.

中文翻译:

OLA生成的囊泡中脂质组成和扩散性的表征。

巨型单层囊泡(GUV)是许多科学分支(包括生物物理学和合成生物学)中的多功能工具。辛醇辅助脂质体组装(OLA)是一种最新开发的微流体技术,能够在高度受控的实验条件下在单个设备中生产和测试GUV。因此,作为用于药物发现,人工细胞生产以及更普遍地用于脂质膜特性的受控研究的平台,它引起了极大的兴趣。在这项工作中,我们通过形成DOPC,DOPG和DOPE的可调二元脂质混合物的GUV来扩展OLA技术的功能。使用光漂白后的荧光恢复,我们研究了脂质在OLA脂质体中的横向扩散系数,并发现所测试的脂质系统的预期值在1μm2/ s的范围内。我们在一系列条件下研究了OLA衍生的GUV,并将结果与​​电铸囊泡进行了比较。总体而言,我们发现用OLA获得的囊泡中脂质的横向扩散系数与通过传统电铸获得的囊泡中脂质的横向扩散系数在数量上相似。我们的结果对OLA衍生的GUV的质量进行了定量的生物物理验证,这将有助于该多功能平台的广泛使用。我们发现用OLA获得的囊泡中脂质的横向扩散系数在数量上类似于通过传统电铸获得的囊泡中脂质的横向扩散系数。我们的结果对OLA衍生的GUV的质量进行了定量的生物物理验证,这将有助于该多功能平台的广泛使用。我们发现用OLA获得的囊泡中脂质的横向扩散系数在数量上类似于通过传统电铸获得的囊泡中脂质的横向扩散系数。我们的结果对OLA衍生的GUV的质量进行了定量的生物物理验证,这将有助于该多功能平台的广泛使用。
更新日期:2020-05-13
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