Protein Encapsulation in Unilamellar Liposomes: High Encapsulation Efficiency and A Novel Technique to Assess Lipid-Protein Interaction
Protein Encapsulation in Unilamellar Liposomes: High Encapsulation Efficiency and A Novel Technique to Assess Lipid-Protein Interaction
复制标题
DOI：
10.1007/s11095-012-0720-x
复制
发表时间：
2012-07-01
影响因子：
3.7
通讯作者：
Burgess, Diane J.
中科院分区：
文献类型：
--
作者：
Xu, Xiaoming;Costa, Antonio;Burgess, Diane J.
为了用一种简单的方法将大量的蛋白质（超氧化物歧化酶（SOD））包封在单层脂质体中，并研究脂质体与蛋白质的相互作用，将预先制备的单层空脂质体与SOD混合并进行冻融循环。为了研究脂质与蛋白质的相互作用，我们采用了一种新的光散射技术，在150 nm左右的波长处可以获得高达50%的蛋白质包封率。冻融循环后，粒度无显著变化。SOD与高浓度胆固醇的DPPC脂质体有较强的相互作用。光散射数据显示，在某些情况下，SOD分子内的脂质bilayer.The方法允许在这里报道的脂质体制备和蛋白质装载操作可以分开，在制造过程中有很大的灵活性。因此，可以制备空脂质体而不考虑蛋白质稳定性，使得制造过程更灵活且易于控制，并最终导致改善的产品质量。为了解释SOD与脂质的相互作用，提出了“口袋嵌入”理论。本文报道的包封方法可以应用于亲水性小分子以及大多数亲水性蛋白质，以实现高包封效率。
To encapsulate a large amount of protein (superoxide dismutase, SOD) into unilamellar liposomes using a simple process and to investigate the lipid-protein interaction.To achieve protein encapsulation, preformed unilamellar empty liposomes were mixed with SOD and subjected to freeze-thaw cycling. To investigate the lipid-protein interaction, a novel light scattering technique was used.Up to 50% protein encapsulation was achieved at similar to 150 nm. There was no significant change in particle size following the freeze-thaw cycling. SOD had a strong interaction with DPPC liposomes containing high concentration of cholesterol. Light scattering data revealed that in some cases the SOD molecules were present inside the lipid bilayer.The method reported here allows great flexibility in the manufacturing process as the liposome preparation and protein-loading operations can be separated. Accordingly, empty liposomes can be prepared without concern about protein stability, making the manufacturing process more flexible and easy to control and ultimately leading to improved product quality. To explain the SOD-lipid interaction, a "pocket-embedding" theory was proposed. The encapsulation method reported here can be applied to hydrophilic small molecules as well as most hydrophilic proteins to achieve high encapsulation efficiency.