Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
Lab Chip. 2012 Feb 7;12(3):562-5. doi: 10.1039/c2lc21129g. Epub 2011 Dec 19.
The development of a method for high-throughput, automated proteomic screening could impact areas ranging from fundamental molecular interactions to the discovery of novel disease markers and therapeutic targets. Surface display techniques allow for efficient handling of large molecular libraries in small volumes. In particular, phage display has emerged as a powerful technology for selecting peptides and proteins with enhanced, target-specific binding affinities. Yet, the process becomes cumbersome and time-consuming when multiple targets are involved. Here we demonstrate for the first time a microfluidic chip capable of identifying high affinity phage-displayed peptides for multiple targets in just a single round and without the need for bacterial infection. The chip is shown to be able to yield well-established control consensus sequences while simultaneously identifying new sequences for clinically important targets. Indeed, the confined parameters of the device allow not only for highly controlled assay conditions but also introduce a significant time-reduction to the phage display process. We anticipate that this easily-fabricated, disposable device has the potential to impact areas ranging from fundamental studies of protein, peptide, and molecular interactions, to applications such as fully automated proteomic screening.
高通量、自动化蛋白质组学筛选方法的发展可能会影响从基本分子相互作用到发现新的疾病标志物和治疗靶点等多个领域。表面展示技术允许在小体积中高效处理大型分子文库。特别是噬菌体展示已成为一种强大的技术,可用于选择具有增强的、针对特定靶标结合亲和力的肽和蛋白质。然而,当涉及多个靶标时,该过程变得繁琐且耗时。在这里,我们首次展示了一种微流控芯片,该芯片能够在单个轮次中识别针对多个靶标的高亲和力噬菌体展示肽,而无需细菌感染。该芯片能够产生成熟的控制共识序列,同时为临床重要靶标识别新序列。事实上,该设备的受限参数不仅允许高度控制的测定条件,而且还将噬菌体展示过程的时间大大缩短。我们预计,这种易于制造的一次性设备具有广泛的应用潜力,从蛋白质、肽和分子相互作用的基础研究到全自动蛋白质组学筛选等应用。