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， 这是多种遗传和蛋白质组改变逐渐积累的结果，也是癌症的原因。 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 科学已经导致了DNA芯片、ELISA、微型生物传感器、微流体装置的发展， (e.g.,生物MEMS或微机电系统和质谱。这些使能技术 已经极大地影响了我们检测和分析癌症的方式，比如基因表达谱， 药物研发和临床诊断。然而，这些技术平台都不够灵活 允许检测遗传改变和蛋白质谱，灵敏度低至单个分子 水平随着当前基因组学和蛋白质组学研究产生更多序列数据，存在强烈需求 用于快速筛选大量核酸和蛋白质的新技术。 在这种情况下，该提案的主要目标是开发一种通用的敏感技术， 以高度多路复用的方式快速分析癌症分子谱（如DNA、RNA和蛋白质）， 准确诊断、诊断和有效治疗的方式。创新和基本原理 该技术的优点在于半导体量子点或QD的新颖光学性质（例如，可调谐 光发射、改善的信号亮度、抗光漂白和同时激发 多种颜色）和我们使用这些纳米颗粒制作光学条形码的能力。不同于单身 基于QD的成像，我们提出制备微米尺寸和纳米尺寸的QD编码的光学条形码。 纳米尺寸。微条形码将用于标记生物分子探针，而纳米球将 可用作报告基因，使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孔密封方法，以稳定光学条形码， 化学缀合方法来优化生物分子探针固定。我们将进一步开展 在DNA和RNA水平上检测突变以及蛋白质癌症谱的实验 生物标志物，其可以从血清或均质化的细胞和组织样本中分离。

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.