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

被引:19
作者
Bright, Leonard K. [1 ]
Baker, Christopher A. [1 ]
Branstrom, Robert [4 ]
Saavedra, S. Scott [1 ,2 ]
Aspinwall, Craig A. [1 ,2 ,3 ]
机构
[1] Univ Arizona, Dept Chem & Biochem, Tucson, AZ 85721 USA
[2] Univ Arizona, Inst BIO5, Tucson, AZ 85721 USA
[3] Univ Arizona, Dept Biomed Engn, Tucson, AZ 85721 USA
[4] Karolinska Inst, Karolinska Univ Hosp, Dept Mol Med & Surg, Endocrine & Sarcoma Surg Unit, SE-17176 Stockholm, Sweden
基金
美国国家卫生研究院;
关键词
black lipid membrane; lipid bilayer; polymer scaffold; methacrylate polymer; ion channel; alpha-hemolysin; gramicidin; PHOSPHOLIPID-VESICLES; MEMBRANE-PROTEIN; FORMING PROTEIN; SILICON CHIPS; GRAMICIDIN; STABILIZATION; PEPTIDES; DNA;
D O I
10.1021/acsbiomaterials.5b00205
中图分类号
TB3 [工程材料学]; R318.08 [生物材料学];
学科分类号
0805 ; 080501 ; 080502 ;
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-functionalized 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 crosslink density. Additionally, the polymer scaffold exerted minimal perturbation to membrane electrical integrity as indicated by mean conductance measurements. When gramicidin A and alpha-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.
引用
收藏
页码:955 / 963
页数:9
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