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Methacrylate Polymer Scaffolding Enhances the Stability of Suspended Lipid Bilayers for Ion Channel Recordings and Biosensor Development

机译:甲基丙烯酸酯聚合物脚手架提高了离子通道记录和生物传感器开发的悬浮脂质双层的稳定性。

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摘要

Black lipid membranes (BLMs) provide a synthetic environment that facilitates measurement of ion channel activity in diverse analytical platforms. The limited electrical, mechanical and temporal stabilities of BLMs pose a significant challenge to development of highly stable measurement platforms. Here, ethylene glycol dimethacrylate (EGDMA) and butyl methacrylate (BMA) were partitioned into BLMs and photopolymerized to create a cross-linked polymer scaffold in the bilayer lamella that dramatically improved BLM stability. The commercially available methacrylate monomers provide a simple, low cost, and broadly accessible approach for preparing highly stabilized BLMs useful for ion channel analytical platforms. When prepared on silane-modified glass microapertures, the resulting polymer scaffold-stabilized (PSS)-BLMs exhibited significantly improved lifetimes of 23 ± 9 to 40 ± 14 h and > 10-fold increase in mechanical stability, with breakdown potentials > 2000 mV attainable, depending on surface modification and polymer cross-link density. Additionally, the polymer scaffold exerted minimal perturbations to membrane electrical integrity as indicated by mean conductance measurements. When gramicidin A and α-hemolysin were reconstituted into PSS-BLMs, the ion channels retained function comparable to conventional BLMs. This approach is a key advance in the formation of stabilized BLMs and should be amenable to a wide range of receptor and ion channel functionalized platforms.
机译:黑色脂质膜(BLM)提供了一个合成环境,可促进在各种分析平台中测量离子通道活性。 BLM有限的电气,机械和时间稳定性对高度稳定的测量平台的开发提出了重大挑战。在这里,将乙二醇二甲基丙烯酸酯(EGDMA)和甲基丙烯酸丁酯(BMA)分配到BLM中并进行光聚合,以在双层薄层中创建交联的聚合物支架,从而显着提高BLM的稳定性。市售的甲基丙烯酸酯单体为制备用于离子通道分析平台的高度稳定的BLM提供了一种简单,低成本且可广泛使用的方法。当在硅烷改性的玻璃微孔上制备时,所得的聚合物支架稳定(PSS)-BLM的使用寿命显着提高,为23±9至40±14 h,机械稳定性提高了> 10倍,击穿电势> 2000 mV取决于表面改性和聚合物交联密度。另外,如平均电导率测量所表明的,聚合物支架对膜的电完整性影响最小。当将短杆菌肽A和α-溶血素重构为PSS-BLM时,离子通道保留的功能与传统BLM相当。这种方法是稳定BLM形成的关键进展,应适用于各种受体和离子通道功能化平台。

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