Ultrasensitive Photoamplifed Fluorescence Detection of Ligand Binding on a Chip

描述（由申请人提供）：分子识别事件的检测一直是生物分析科学的主要关注领域，因为细胞中生物分子对象与其他分子实体之间的结合在分子水平上控制所有细胞功能。检测生物分子之间相互作用的能力对于开发新疗法和治疗剂至关重要。该提案的目标是开发一种全新的方法，用于生物分子识别事件的超灵敏放大检测，并将其应用于低成本的生物分析微阵列芯片。所提出的方法-这是基于电子转移敏化剂的自放大解蔽-是标准化的和通用的，即不限于选定类别的生物分子和相互作用。一旦使用已知的配体-受体对进行验证，该技术将应用于分子生物学中的一个突出问题：识别组蛋白尾部的翻译后修饰。在染色质领域的一个至关重要的问题是确定新的结构域能够阅读组蛋白密码。鉴于组蛋白尾部存在大量的翻译后修饰，主要来源于赖氨酸和精氨酸残基的甲基化或乙酰化，因此只能通过高通量组合筛选来寻找新的阅读器结构域。将合成含有天然组蛋白尾部的特征残基的七肽文库，将其打印在扩增检测微阵列芯片上，并筛选已知“读取”组蛋白代码的结构域-染色体结构域、溴结构域和PHD指的结合。 公共卫生相关性检测生物分子之间的相互作用一直是生物医学科学的主要关注领域，因为这种相互作用控制着活生物体中的所有过程。检测生物分子之间结合的能力对于开发新疗法和治疗剂至关重要。该提案的目标是开发一种全新的低成本超灵敏检测方法，这将有助于发现新药。

DESCRIPTION (provided by applicant): Detection of molecular recognition events has always been an area of primary focus in bioanalytical sciences, as binding between biomolecular objects and other molecular entities in cells governs all cellular functions at the molecular level. The ability to detect interactions between biological molecules is vitally important for the development of new therapies and therapeutic agents. The goal of this proposal is to develop a fundamentally new approach to ultra-sensitive amplified detection of biomolecular recognition events and implement it in low cost bioanalytical microarray chips. The proposed methodology - which is based on self-amplified unmasking of electron-transfer sensitizers - is standardized and universal, i.e. not limited to selected classes of biological molecules and interactions. Once validated using known ligand-receptor pairs, this technology will be applied to a prominent problem in molecular biology: recognition of posttranslational modifications of histone tails. A critically important problem in the chromatin field is to identify new domains capable of reading the histone code. Given a large number of posttranslational modifications in histone tails, derived primarily from methylation or acetylation of their lysine and arginine residues, the search for new reader domains can only be achieved with high throughput combinatorial screening. Heptapeptide libraries containing signature residues of natural histone tails will be synthesized, printed on the amplified detection microarray chips, and screened for binding of domains known to "read" the histone code - chromodomain, bromodomain and the PHD finger. PUBLIC HEALTH RELEVANCE Detection of interactions between biological molecules has always been an area of primary focus in biomedical sciences, as such interactions govern all processes in living organisms. The ability to detect binding between biological molecules is vitally important for the development of new therapies and therapeutic agents. The goal of this proposal is to develop a fundamentally new approach to low cost ultra-sensitive detection methods, which will aid in the discovery of new medicines.