Laboratory of Nanostructures and Biosensing, Department of Electrical and Computer Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA.
ACS Nano. 2011 Sep 27;5(9):7555-64. doi: 10.1021/nn202554t. Epub 2011 Aug 17.
Microarray technology has facilitated many powerful high-throughput studies in the fields of genetics and proteomics, among others. However, preparation of microarrays composed of cell-derived membranes with embedded receptors has proven difficult. Here we describe a new method for forming microarrays composed of synthetic lipid vesicles and natural cell membranes. The method is based upon assembly of vesicles and natural membranes into recessed micro- and nanowells and using a polydimethylsiloxane (PDMS) block as a "squeegee." This method is used to assemble phospholipid vesicles into arrays with micrometer and nanoscale dimensions. Native myelin and neuronal lipid raft arrays are also formed in 30 min or less. We show the natural membrane arrays can be used for sensing lipid-protein interactions by detecting cholera toxin binding to ganglioside GM1 in neuronal lipid rafts. In multicomponent arrays myelin can be distinguished from neuronal rafts by antibody binding to cell-specific surface antigens. Finally, myelin arrays formed in gold nanowells are used for surface plasmon resonance sensing. This assembly approach is simple, broadly applicable, and opens up new avenues of research not easily accomplished with standard microarray technology.
微阵列技术促进了遗传学和蛋白质组学等领域的许多强大的高通量研究。然而，制备含有嵌入式受体的细胞衍生膜的微阵列一直很困难。在这里，我们描述了一种新的方法，用于形成由合成脂质体和天然细胞膜组成的微阵列。该方法基于将囊泡和天然膜组装到凹陷的微纳米井中，并使用聚二甲基硅氧烷 (PDMS) 块作为“刮刀”。该方法用于将磷脂囊泡组装成具有微米和纳米尺寸的阵列。天然髓鞘和神经元脂筏阵列也可以在 30 分钟或更短的时间内形成。我们展示了天然膜阵列可用于通过检测神经脂筏中的神经节苷脂 GM1 与霍乱毒素的结合来检测脂质-蛋白质相互作用。在多组分阵列中，通过针对细胞特异性表面抗原的抗体结合，可以将髓鞘与神经元筏区分开来。最后，使用金纳米井中的髓鞘阵列进行表面等离子体共振感测。这种组装方法简单、应用广泛，并为使用标准微阵列技术难以完成的新研究开辟了途径。
