Mayer L D, Hope M J, Cullis P R
Biochim Biophys Acta. 1986 Jun 13;858(1):161-8. doi: 10.1016/0005-2736(86)90302-0.
Previous studies from this laboratory have shown that large unilamellar vesicles can be efficiently produced by extrusion of multilamellar vesicles through polycarbonate filters with a pore size of 100 nm (Hope, M.J., Bally, M.B., Webb, G. and Cullis, P.R. (1985) Biochim. Biophys. Acta 812, 55-65). In this work it is shown that similar procedures can be employed for the production of homogeneously sized unilamellar or plurilamellar vesicles by utilizing filters with pore sizes ranging from 30 to 400 nm. The unilamellarity and trapping efficiencies of these vesicles can be significantly enhanced by freezing and thawing the multilamellar vesicles prior to extrusion. This procedure is particularly applicable when very high lipid concentrations (400 mg/ml) are used, where extrusion of the frozen and thawed multilamellar vesicles through 100 and 400 nm filters results in trapping efficiencies of 56 and 80%, respectively. Freeze-fracture electron microscopy revealed that vesicles produced at these lipid concentrations exhibit size distributions and extent of multilamellar character comparable to systems produced at lower lipid levels. These results indicate that the freeze-thaw and extrusion process is the technique of choice for the production of vesicles of variable sizes and high trapping efficiency.
该实验室之前的研究表明,通过将多层囊泡挤压通过孔径为100 nm的聚碳酸酯滤膜,可以高效地制备大单层囊泡(Hope, M.J., Bally, M.B., Webb, G. 和Cullis, P.R. (1985) Biochim. Biophys. Acta 812, 55 - 65)。在这项工作中表明,通过使用孔径范围为30至400 nm的滤膜,类似的方法可用于制备尺寸均匀的单层或多层囊泡。在挤压之前对多层囊泡进行冻融处理,可以显著提高这些囊泡的单层性和包封效率。当使用非常高的脂质浓度(400 mg/ml)时,该方法特别适用,此时将冻融后的多层囊泡通过100 nm和400 nm滤膜挤压,包封效率分别为56%和80%。冷冻断裂电子显微镜显示,在这些脂质浓度下产生的囊泡,其尺寸分布和多层特征程度与在较低脂质水平下产生的系统相当。这些结果表明,冻融和挤压过程是制备不同尺寸且包封效率高的囊泡的首选技术。