在高胆固醇含量、带电脂质和生理盐水溶液条件下,湿膜中巨型单拉美米尔囊泡的电形成。

IF 3.3 4区 工程技术 Q2 CHEMISTRY, PHYSICAL Membranes Pub Date : 2024-10-12 DOI:10.3390/membranes14100215
Ivan Mardešić, Zvonimir Boban, Marija Raguz
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引用次数: 0

摘要

在膜特性研究中,巨型单淀粉囊泡经常被用作膜模型。它们最常用的是电形成法。然而,有许多参数会影响该过程的成功与否。已证明会对 GUV 电形成产生负面影响的一些最常见的条件包括胆固醇(Chol)浓度过高、使用含有带电脂质的混合物以及离子强度过高的溶液。高浓度胆固醇对传统的电铸工艺来说是个问题,因为它需要通过完全蒸发脂质混合物中的有机溶剂来形成干燥的脂质膜。在干燥过程中,会形成无水胆醇晶体。它们不参与脂质双分子层的形成,因此与原始脂质混合物相比,囊泡双分子层中的胆醇浓度较低。我们主要是受人造胆醇脱混问题的启发,对电铸方案进行了修改,将快速溶剂交换(RSE)、超声波处理、等离子清洗和旋涂等技术结合在一起,以便从潮湿的脂质薄膜中可重复地生产出 GUV。除了减少 Chol 脱混现象,我们还证明该方法还可用于从带电脂质混合物中以及在用作内部溶液的离子溶液中生产 GUV。在混合比为 0 至 2.5 的 Chol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) 样品中,我们获得了高产率的 GUV。我们还成功地在低离子强度的氯化钠溶液中从含有高达 60 摩尔%的带电荷脂质 1-棕榈酰-2-油酰基-正-甘油-3-磷-L-丝氨酸(POPS)的混合物中制备出了 GUV。
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Electroformation of Giant Unilamellar Vesicles from Damp Films in Conditions Involving High Cholesterol Contents, Charged Lipids, and Saline Solutions.

Giant unilamellar vesicles (GUVs) are frequently used as membrane models in studies of membrane properties. They are most often produced using the electroformation method. However, there are a number of parameters that can influence the success of the procedure. Some of the most common conditions that have been shown to have a negative effect on GUV electroformation are the presence of high cholesterol (Chol) concentrations, the use of mixtures containing charged lipids, and the solutions with an elevated ionic strength. High Chol concentrations are problematic for the traditional electroformation protocol as it involves the formation of a dry lipid film by complete evaporation of the organic solvent from the lipid mixture. During drying, anhydrous Chol crystals form. They are not involved in the formation of the lipid bilayer, resulting in a lower Chol concentration in the vesicle bilayer compared to the original lipid mixture. Motivated primarily by the issue of artifactual Chol demixing, we have modified the electroformation protocol by incorporating the techniques of rapid solvent exchange (RSE), ultrasonication, plasma cleaning, and spin-coating for reproducible production of GUVs from damp lipid films. Aside from decreasing Chol demixing, we have shown that the method can also be used to produce GUVs from lipid mixtures with charged lipids and in ionic solutions used as internal solutions. A high yield of GUVs was obtained for Chol/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) samples with mixing ratios ranging from 0 to 2.5. We also succeeded in preparing GUVs from mixtures containing up to 60 mol% of the charged lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) and in NaCl solutions with low ionic strength (<25 mM).

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来源期刊
Membranes
Membranes Chemical Engineering-Filtration and Separation
CiteScore
6.10
自引率
16.70%
发文量
1071
审稿时长
11 weeks
期刊介绍: Membranes (ISSN 2077-0375) is an international, peer-reviewed open access journal of separation science and technology. It publishes reviews, research articles, communications and technical notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided.
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