Surface Immobilization of Bio-Functionalized Cubosomes: Sensing of Proteins by Quartz Crystal Microbalance
Surface Immobilization of Bio-Functionalized Cubosomes: Sensing of Proteins by Quartz Crystal Microbalance
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DOI：
10.1021/la2032994
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发表时间：
2012-01-10
期刊：
影响因子：
3.9
通讯作者：
Separovic, Frances
中科院分区：
文献类型：
--
作者：
Fraser, Scott J.;Mulet, Xavier;Separovic, Frances
报道了一种将脂质液晶亚微米颗粒（立方体）拴在金表面用于蛋白质检测的策略。采用时间分辨石英晶体微天平（QCM-D）对蛋白质与立方体的相互作用进行了真实的实时监测。为了实现特异性结合，从非离子表面活性剂植烷三醇、嵌段共聚物、Pluronic F-127和二级生物素化脂质、1，2-二硬脂酰-sn-甘油基-3-磷酸乙醇胺N-[生物素基（聚乙二醇）-2000]制备立方体，其使得颗粒能够附着到QCM传感器芯片的金表面处的中性抗生物素蛋白（NAv）-烷醇单分子层。通过添加糖脂（G（M1））进一步官能化第二组立方体，以促进蛋白质霍乱毒素B亚基（CT B）从溶液中的特异性结合摄取。QCM-D证实了立方体-NAv结合的特异性。同样使用QCM进行的滴定实验分析表明，颗粒在表面的有效包装需要最佳浓度的立方体：高立方体浓度导致立方体混乱地结合到表面上，空间抑制表面附着，或需要显着重组以允许均匀的立方体覆盖。该方法能够直接制备复杂的纳米结构的edibrium，然后用于从溶液中特异性捕获分析物蛋白（霍乱毒素B亚基或游离NAV），支持这种方法作为生物传感平台的发展潜力。
A strategy for tethering lipid liquid crystalline submicrometer particles (cubosomes) to a gold surface for the detection of proteins is reported. Time-resolved quartz crystal microbalance (QCM-D) was used to monitor the cubosome-protein interaction in real time. To achieve specific binding, cubosomes were prepared from the nonionic surfactant phytantriol, block-copolymer, Pluronic F-127, and a secondary biotinylated lipid, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine N-[biotinyl(polyethyleneglycol)-2000], which enabled attachment of the particles to a neutravidin (NAv)-alkanethiol monolayer at the gold surface of the QCM sensor chip. A second set of cubosomes was further functionalized with addition of the glycolipid (G(M1)) to facilitate a specific binding uptake of the protein, cholera toxin B subunit (CTB), from solution. QCM-D confirmed the specificity of the cubosome-NAv binding. The analysis of titration experiments, also performed with QCM, suggests that an optimal concentration of cubosomes is required for the efficient packing of the particles at the surface: high cubosome concentrations lead to chaotic cubosome binding onto the surface, sterically inhibiting surface attachment, or require significant reorganization to permit uniform cubosome coverage. The methodology enabled the straightforward preparation of a complex nanostructured edifice, which was then used to specifically capture analyte proteins (cholera toxin B subunit or free NAv) from solution, supporting the potential for development of this approach as a biosensing platform.