文献：聚乙二醇负载脂质双层在金表面的原位形成及表征

链接：https://pubs.acs.org/doi/abs/10.1021/la048378o

作者：杰弗里·C·芒罗，柯蒂斯·W·弗兰克

摘要：

有人提出，在脂质双层膜和固体表面之间添加聚合物垫层，可以形成柔软、可变形的膜层，从而实现跨膜蛋白的插入和迁移。本研究通过两步吸附过程，在聚乙二醇 (PEG) 载体上形成了可迁移的、可束缚的脂质双层膜。 PEG 薄膜是通过共吸附异功能远爪 PEG 脂质聚合物（1,2-二硬脂酰-sn-甘油-3-磷酸乙醇胺-N-聚（乙二醇）-2000- N- [3-（2-（吡啶基二硫代）丙酸酯]）（DSPE-PEG-PDP）和非脂质功能化的 PEG-PDP 从乙醇/水混合物中制备的，如先前的论文（Munro，JC；Frank，CW Langmuir 2004，20，3339-3349）中所述。

然后使用两步脂质吸附策略。首先，将脂质从己烷溶液中吸附到 PEG 载体上。其次，将囊泡吸附并融合在表面以在水环境中形成双层。光漂白实验后的荧光恢复表明，该过程产生扩散系数约为 2 μm /s。随着束缚脂质密度的增加，双层膜的迁移率略有降低。表面等离子体共振法（用于测定原位膜厚度）和荧光法（用于定量测定每个18 x 18毫米样品的荧光强度）也证实了双层膜的形成，而非多层结构。遗憾的是，荧光显微镜检查也显示样品上存在较大的缺陷，这限制了该系统的实用性。

Abstract Image

Inclusion of a polymer cushion between a lipid bilayer membrane and a solid surface has been suggested as a means to provide a soft, deformable layer that will allow for transmembrane protein insertion and mobility. In this study, mobile, tethered lipid bilayers were formed on a poly(ethylene glycol) (PEG) support via a two-step adsorption process. The PEG films were prepared by coadsorbing a heterofunctional, telechelic PEG lipopolymer (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-poly(ethylene glycol)-2000-N-[3-(2-(pyridyldithio)propionate]) (DSPE-PEG-PDP) and a nonlipid functionalized PEG-PDP from an ethanol/water mixture, as described in a previous paper (Munro, J. C.; Frank, C. W. Langmuir2004, 20, 3339−3349). Then a two-step lipid adsorption strategy was used. First, lipids were adsorbed onto the PEG support from a hexane solution. Second, vesicles were adsorbed and fused on the surface to create a bilayer in an aqueous environment. Fluorescence recovery after photobleaching experiments show that this process results in mobile bilayers with diffusion coefficients on the order of 2 μm2/s. The mobility of the bilayers is decreased slightly by increasing the density of tethered lipids. The formation of bilayers, and not multilayer structures, is also confirmed by surface plasmon resonance, which was used to determine in situ film thickness, and by fluorimetry, which was used to determine quantitatively the fluorescence intensity for each 18 by 18 mm sample. Unfortunately, fluorescence microscopy also shows that there are large defects on the samples, which limits the utility of this system.

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