Sensitive and multiplexed analysis of cancer biomarkers with QD barcodes

癌症被认为是一种涉及无数分子过程的高度复杂的疾病，其产生的原因是 多种基因和蛋白质组改变逐渐积累的结果，这些改变也可用作癌症 生物标志物。当前癌症研究的一个主要焦点是如何将这些潜在的分子事件联系起来 与癌症的发展和进展有关。肿瘤分子分析和生物分析的最新进展 科学导致了DNA芯片、酶联免疫吸附试验、微型生物传感器、微流控设备的发展 (例如，生物MEMS或微电子机械系统和质谱仪)。这些使能技术 在很大程度上影响了我们检测和分析癌症的方式，例如基因表达谱， 药物发现和临床诊断学。然而，这些技术平台都不够灵活 允许检测基因改变和蛋白质图谱，灵敏度可低至单分子 水平。随着目前基因组学和蛋白质组学的研究产生更多的序列数据，存在着强烈的需求 用于能够快速筛选大量核酸和蛋白质的新技术。 在这种情况下，这项提议的主要目标是开发一种通用和敏感的技术，能够 快速分析高度多元化的癌症分子图谱(如DNA、RNA和蛋白质) 提供准确的诊断、预后和有效的治疗方法。创新与基本理念 这项技术的关键在于半导体量子点或量子点(例如，可调谐的)的新颖光学性质 发光、改善信号亮度、抗光漂白和同时激发 多种颜色)以及我们使用这些纳米颗粒制造光学条形码的能力。不同于单身 基于量子点的成像，我们建议制备微米级和微米级的量子点编码光学条形码 纳米尺寸。微型条形码将被用来标记生物分子探测器，而纳米球将被用来标记 作为记录器，将电流检测灵敏度提高2-3个数量级。我们还将 探索新的表面封装和气孔密封方法来稳定光学条形码，以及 优化生物分子探针固定化的化学共轭方法。我们将进一步开展 检测DNA和RNA水平上的突变以及蛋白质癌特征的实验 生物标志物，可以从血清或匀浆的细胞和组织标本中分离出来。

Cancer is recognized as a highly complex disease involving myriad molecular processes and arises as the result of gradual accumulation of multiple genetic and proteomic alterations, which also serve as cancer biomarkers. A major focus of current cancer research is how to correlate these underlying molecular events with cancer development and progression. Recent advances in cancer molecular analysis and bioanalytical sciences have led to the development of DNA chips, ELISA, miniaturized biosensors, microfluidic devices (e.g., bioMEMS or microelectromechanical systems, and mass spectrometry. These enabling technologies have substantially influenced the way that we detect and analyze cancer, such as gene expression profiling, drug discovery, and clinical diagnostics. However, none of these technical platforms are sufficiently flexible to allow detection of both genetic alterations and protein profiles with sensitivity down to single molecule level. As current research in genomics and proteomics produces more sequence data, there is a strong need for new technologies that can rapidly screen a large number of nucleic acids and proteins. In this context, the primary goal of this proposal is to develop a versatile and sensitive technology that can quickly analyze cancer molecular profiles (such as DNAs, RNAs and proteins) in a highly multiplexed manner for accurate diagnostics, prognostics and effective therapeutics. The innovation and basic rationale of this technology lies in the novel optical properties of semiconductor quantum dots or QDs (e.g., tunable light emission, improved signal brightness, resistance against photobleaching, and simultaneous excitation of multiple colors) and our ability to make optical barcodes using these nanoparticles. Different from single QD based imaging, we propose to prepare QD encoded optical barcodes of both micrometer sizes and nanometer sizes. The micro-barcodes will be used to tag biomolecular probes, whereas the nanospheres will be used as reporters to enhance the current detection sensitivity by 2-3 orders of magnitude. We will also explore new surface encapsulation and pore sealing approaches to stabilize the optical barcodes, and chemical conjugation approaches to optimize biomolecular probe immobilization. We will further carry out experiments to detect mutations on both DNA and RNA level as well as the profiles of protein cancer biomarkers, which can be isolated from serum or homogenized cell and tissue specimens.