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Enhanced avidin binding to lipid bilayers using PDP-PE lipids with PEG-biotin linkers
Nanoscale Advances ( IF 4.6 ) Pub Date : 2020-03-10 , DOI: 10.1039/d0na00060d
Holly L Birchenough 1 , Marcus J Swann 2 , Egor Zindy 1 , Anthony J Day 1 , Thomas A Jowitt 1
Affiliation  

Two of the most important aspects of lipid bilayers that have increased their popularity in the field of nanotechnology and biosensors are their fluid nature, which is highly beneficial in ensuring the spatial organization of attached molecules, and the relative ease in which they can be manipulated to change the surface chemistry. Here we have used two different types of functionalized lipids to study the interaction of avidin, which is a common approach to attach further ligands for study. We have tested the commonly used Biotinyl-Cap-PE lipids at different molar percentages and reveal that avidin is not evenly distributed, but forms what looks like clusters even at low percentage occupancy which hampers the level of avidin that can be associated with the surface. We have then successfully employed the novel strategy of using PDP-PE lipids which contain a reducible disulphide to which we added maleamide-PEG-biotin spacers of different lengths. There is a more even distribution of avidin on these layers and thereby increasing the amount and efficiency of avidin association. The reduced levels of avidin that was being associated with the Biotinyl-Cap-PE layers as compared to the PDP-PE lipids could be analysed with QCM-D and interferometry approaches, but it was only with SEEC microscopy that the reason for the reduced occupancy was resolved.

中文翻译:

使用具有 PE​​G-生物素接头的 PDP-PE 脂质增强抗生物素蛋白与脂质双层的结合

脂质双层在纳米技术和生物传感器领域越来越受欢迎的两个最重要的方面是它们的流体性质,这对于确保附着分子的空间组织非常有益,以及它们可以相对容易地被操纵到改变表面化学。在这里,我们使用了两种不同类型的功能化脂质来研究抗生物素蛋白的相互作用,这是附加更多配体进行研究的常用方法。我们以不同的摩尔百分比测试了常用的 Biotinyl-Cap-PE 脂质,发现抗生物素蛋白分布不均匀,但即使在低占有率的情况下也会形成看起来像簇的物质,这会阻碍与表面相关的抗生物素蛋白水平。然后,我们成功地采用了使用含有可还原二硫化物的 PDP-PE 脂质的新策略,我们在其中添加了不同长度的马来酰胺-PEG-生物素间隔物。在这些层上有更均匀的抗生物素蛋白分布,从而增加了抗生物素蛋白结合的数量和效率。与 PDP-PE 脂质相比,与 Biotinyl-Cap-PE 层相关的抗生物素蛋白水平降低可以使用 QCM-D 和干涉测量方法进行分析,但只有使用 SEEC 显微镜才能分析占用率降低的原因已解决。
更新日期:2020-04-24
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