利用薄膜挤压法制备支撑脂双层膜
发布时间：2018-07-25 12:40
【摘要】：背景:人工支撑脂双层膜是最接近细胞膜的体外仿生模型,构建结构与功能均类似于真实细胞膜的脂双层膜具有十分重要的意义。目的:探索表面均匀且流动性高的支撑脂双层膜的最佳制备条件并检测指标。方法:采用薄膜挤压法,分别以孔径为0.1,0.2μm的滤膜对脂质体分子过滤10次,保留未过滤样品作为对照组,通过原子力显微镜对其表面粗糙程度进行检测,探讨制备均匀程度较高的脂双层膜所需滤膜的最佳孔径。将160μL卵磷脂(10 g/L)分别与5,10,25μL荧光磷脂(1 g/L)进行混合,通过荧光漂白恢复技术对脂双层膜的流动性进行检测,探讨卵磷脂与荧光磷脂的最适比例。结果与结论:原子力显微镜实验结果显示,经过孔径为0.1μm滤膜过滤所得的脂双层膜表面均匀程度最高(P0.01),其表面均方根粗糙度为(0.432±0.181)nm,当滤膜孔径大于0.1μm时,脂双层膜表面均匀程度差且存在较多的未铺展开脂质体囊泡。荧光漂白恢复实验结果显示,采用孔径为0.1μm滤膜过滤的脂双层膜荧光恢复程度最高,当卵磷脂(10 g/L)与荧光磷脂(1 g/L)体积比为160∶10时,采用孔径为0.1μm滤膜过滤的脂双层膜荧光恢复程度达到90%,且扩散系数大于1μm~2/s,符合优质脂双层膜的标准。实验提示利用薄膜挤压法制备脂双层膜时,卵磷脂与荧光磷脂的最适比例以及最佳滤膜孔径。
[Abstract]:Background: artificially supported lipid bilayer membrane is the closest biomimetic model in vitro. It is very important to construct lipid bilayer membrane with structure and function similar to that of real cell membrane. Objective: to explore the optimum preparation conditions and detection index of the lipid bilayer membrane with uniform surface and high fluidity. Methods: the surface roughness of liposomes was determined by atomic force microscope (AFM). The liposome molecules were filtered 10 times by the filter membrane with the pore diameter of 0.1 渭 m and the unfiltered samples were retained as the control group by the thin film extrusion method, and the surface roughness of the liposomes was determined by atomic force microscope (AFM). The optimum pore size of lipid bilayer membrane with high homogeneity was discussed. 160 渭 L lecithin (10 g / L) was mixed with 5 10 渭 L fluorescent phospholipid (1 g / L). The fluidity of lipid bilayer membrane was detected by fluorescence bleaching recovery technique, and the optimum ratio of lecithin to fluorescent phospholipid was discussed. Results and conclusion: the results of atomic force microscope (AFM) showed that the surface uniformity of lipid bilayer membrane was the highest (P0.01), and the RMS roughness was (0.432 卤0.181) nm. When the pore diameter of the membrane was greater than 0.1 渭 m, the surface homogeneity of lipid bilayer membrane was the highest (P0.01), and the mean square roughness of the membrane was (0.432 卤0.181) nm. The surface uniformity of lipid bilayer membrane was poor and there were more unspread liposome vesicles. The results of fluorescence bleaching recovery experiment showed that the fluorescence recovery of lipid bilayer membrane was the highest when the volume ratio of lecithin (10 g / L) to fluorescent phospholipid (1 g / L) was 160: 10. The fluorescence recovery degree of lipid bilayer membrane filtered with 0.1 渭 m pore diameter was 90%, and the diffusion coefficient was more than 1 渭 m ~ 2 / s, which met the standard of high quality lipid bilayer membrane. It is suggested that the optimum ratio of lecithin to fluorescent phospholipids and the optimum pore size of the membrane can be obtained when the bilayer lipids are prepared by thin-film extrusion.
【作者单位】： 华南理工大学生物科学与工程学院;
【基金】：国家自然科学基金(11672109,11272125,11432006)~~
【分类号】：R318.08
，
本文编号：2143856
[Abstract]:Background: artificially supported lipid bilayer membrane is the closest biomimetic model in vitro. It is very important to construct lipid bilayer membrane with structure and function similar to that of real cell membrane. Objective: to explore the optimum preparation conditions and detection index of the lipid bilayer membrane with uniform surface and high fluidity. Methods: the surface roughness of liposomes was determined by atomic force microscope (AFM). The liposome molecules were filtered 10 times by the filter membrane with the pore diameter of 0.1 渭 m and the unfiltered samples were retained as the control group by the thin film extrusion method, and the surface roughness of the liposomes was determined by atomic force microscope (AFM). The optimum pore size of lipid bilayer membrane with high homogeneity was discussed. 160 渭 L lecithin (10 g / L) was mixed with 5 10 渭 L fluorescent phospholipid (1 g / L). The fluidity of lipid bilayer membrane was detected by fluorescence bleaching recovery technique, and the optimum ratio of lecithin to fluorescent phospholipid was discussed. Results and conclusion: the results of atomic force microscope (AFM) showed that the surface uniformity of lipid bilayer membrane was the highest (P0.01), and the RMS roughness was (0.432 卤0.181) nm. When the pore diameter of the membrane was greater than 0.1 渭 m, the surface homogeneity of lipid bilayer membrane was the highest (P0.01), and the mean square roughness of the membrane was (0.432 卤0.181) nm. The surface uniformity of lipid bilayer membrane was poor and there were more unspread liposome vesicles. The results of fluorescence bleaching recovery experiment showed that the fluorescence recovery of lipid bilayer membrane was the highest when the volume ratio of lecithin (10 g / L) to fluorescent phospholipid (1 g / L) was 160: 10. The fluorescence recovery degree of lipid bilayer membrane filtered with 0.1 渭 m pore diameter was 90%, and the diffusion coefficient was more than 1 渭 m ~ 2 / s, which met the standard of high quality lipid bilayer membrane. It is suggested that the optimum ratio of lecithin to fluorescent phospholipids and the optimum pore size of the membrane can be obtained when the bilayer lipids are prepared by thin-film extrusion.
【作者单位】： 华南理工大学生物科学与工程学院;
【基金】：国家自然科学基金(11672109,11272125,11432006)~~
【分类号】：R318.08
，
本文编号：2143856
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