Rieder A A, Koller D, Lohner K, Pabst G
University of Graz, Institute of Molecular Biosciences, Biophysics Division, NAWI Graz, Humboldtstr. 50/III, A-8010 Graz, Austria; BioTechMed-Graz, Austria.
University of Graz, Institute of Molecular Biosciences, Biophysics Division, NAWI Graz, Humboldtstr. 50/III, A-8010 Graz, Austria; BioTechMed-Graz, Austria.
Chem Phys Lipids. 2015 Feb;186:39-44. doi: 10.1016/j.chemphyslip.2014.12.001. Epub 2014 Dec 20.
We have enhanced the rapid solvent exchange (RSE) apparatus by adding controls in temperature, evacuation speed and vortex velocity. Following published protocols yielded vesicles of diverse size and lamellarity as detected by differential scanning calorimetry, photon correlation spectroscopy and X-ray experiments. To optimize the net production of multilamellar vesicles (MLVs) we varied in addition to vortex and evacuation speed lipid and organic solvent concentration, as well as composition of the aqueous medium. Reducing vortexing frequencies and speed of degassing were most beneficial for the yield in MLVs. Additionally also high lipid concentrations and organic solvent/buffer ratios supported MLV formation. To explain our findings we hypothesize on the role of microscopic instabilities on the aqueous phase, which may act as molds for vesicle formation.
我们通过增加温度、抽气速度和涡旋速度的控制,改进了快速溶剂交换(RSE)装置。按照已发表的方案制备出的囊泡,经差示扫描量热法、光子相关光谱法和X射线实验检测,具有不同的大小和片层结构。为了优化多层囊泡(MLV)的净产量,除了涡旋和抽气速度外,我们还改变了脂质和有机溶剂的浓度以及水相介质的组成。降低涡旋频率和脱气速度对MLV的产量最为有利。此外,高脂质浓度和有机溶剂/缓冲液比例也有助于MLV的形成。为了解释我们的发现,我们推测了水相微观不稳定性的作用,它可能充当囊泡形成的模板。