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
{"title":"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。","authors":"Ivan Mardešić, Zvonimir Boban, Marija Raguz","doi":"10.3390/membranes14100215","DOIUrl":null,"url":null,"abstract":"<p><p>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).</p>","PeriodicalId":18410,"journal":{"name":"Membranes","volume":"14 10","pages":""},"PeriodicalIF":3.3000,"publicationDate":"2024-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510074/pdf/","citationCount":"0","resultStr":"{\"title\":\"Electroformation of Giant Unilamellar Vesicles from Damp Films in Conditions Involving High Cholesterol Contents, Charged Lipids, and Saline Solutions.\",\"authors\":\"Ivan Mardešić, Zvonimir Boban, Marija Raguz\",\"doi\":\"10.3390/membranes14100215\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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).</p>\",\"PeriodicalId\":18410,\"journal\":{\"name\":\"Membranes\",\"volume\":\"14 10\",\"pages\":\"\"},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-10-12\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510074/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Membranes\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.3390/membranes14100215\",\"RegionNum\":4,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, PHYSICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Membranes","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.3390/membranes14100215","RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, PHYSICAL","Score":null,"Total":0}
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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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).

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
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.
期刊最新文献
The Application of TiO2/ZrO2-Modified Nanocomposite PES Membrane for Improved Permeability of Textile Dye in Water. Computational Fluid Dynamics Modelling of Hydrogen Production via Water Splitting in Oxygen Membrane Reactors. Fouling of Reverse Osmosis (RO) and Nanofiltration (NF) Membranes by Low Molecular Weight Organic Compounds (LMWOCs), Part 1: Fundamentals and Mechanism. The Influence of Cholesterol on Membrane Targeted Bioactive Peptides: Modulating Peptide Activity Through Changes in Bilayer Biophysical Properties. Cell Type-Specific Anti- and Pro-Oxidative Effects of Punica granatum L. Ellagitannins.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1