Department of Physical and Analytical Chemistry, BMC, Uppsala University, SE-751 23 Uppsala, Sweden.
Anal Biochem. 2010 Oct 15;405(2):153-9. doi: 10.1016/j.ab.2010.06.030. Epub 2010 Jun 19.
The aim of this study was to investigate the potential of polyethylene glycol (PEG)-stabilized lipid bilayer disks as model membranes for surface plasmon resonance (SPR)-based biosensor analyses. Nanosized bilayer disks that included 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)(2000)] (DSPE-PEG(2000)-biotin) were prepared and structurally characterized by cryo-transmission electron microscopy (cryo-TEM) imaging. The biotinylated disks were immobilized via streptavidin to three different types of sensor chips (CM3, CM4, and CM5) varying in their degree of carboxymethylation and thickness of the dextran matrix. The bilayer disks were found to interact with and bind stably to the streptavidin-coated sensor surfaces. As a first step toward the use of these bilayer disks as model membranes in SPR-based studies of membrane proteins, initial investigations were carried out with cyclooxygenases 1 and 2 (COX 1 and COX 2). Bilayer disks were preincubated with the respective protein and thereafter allowed to interact with the sensor surface. The signal resulting from the interaction was, in both cases, significantly enhanced as compared with the signal obtained when disks alone were injected over the surface. The results of the study suggest that bilayer disks constitute a new and promising type of model membranes for SPR-based biosensor studies.
本研究旨在探讨聚乙二醇（PEG）稳定的脂质双层盘作为基于表面等离子体共振（SPR）的生物传感器分析的模型膜的潜力。制备了包含 1,2-二硬脂酰-sn-甘油-3-磷酸乙醇胺-N-[生物素化（聚乙二醇）（2000）]（DSPE-PEG（2000）-生物素）的纳米级双层盘，并通过冷冻传输电子显微镜（cryo-TEM）成像对其进行了结构表征。生物素化的双层盘通过链霉亲和素固定在三种不同类型的传感器芯片（CM3、CM4 和 CM5）上，这些传感器芯片在羧甲基化程度和葡聚糖基质的厚度上有所不同。发现双层盘与链霉亲和素涂层的传感器表面相互作用并稳定结合。作为将这些双层盘用作 SPR 研究膜蛋白的模型膜的第一步，首先对环氧合酶 1 和 2（COX 1 和 COX 2）进行了初步研究。将双层盘与各自的蛋白质预孵育，然后允许其与传感器表面相互作用。在这两种情况下，与单独将双层盘注入表面时相比，相互作用产生的信号都显著增强。研究结果表明，双层盘构成了基于 SPR 的生物传感器研究中一种新型且有前途的模型膜。
