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中主要关注的领域，因为生物分子物体与细胞中其他分子实体之间的结合在分子水平上控制着所有细胞功能。检测生物分子之间相互作用的能力对于新疗法和治疗剂的开发至关重要。本提案的目标是开发一种全新的方法来超灵敏放大检测生物分子识别事件，并在低成本的生物分析微阵列芯片中实现它。所提出的方法-基于电子转移敏化剂的自我放大揭露-是标准化和通用的，即不限于选定的生物分子类别和相互作用。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：组蛋白尾部翻译后修饰的识别。染色质领域的一个关键问题是确定能够读取组蛋白密码的新结构域。考虑到组蛋白尾部大量的翻译后修饰，主要来源于赖氨酸和精氨酸残基的甲基化或乙酰化，寻找新的解读域只能通过高通量组合筛选来实现。合成含有天然组蛋白尾部特征残基的七肽文库，将其打印在扩增的检测芯片上，并筛选已知的“读取”组蛋白编码域的结合-色域，溴域和PHD指。检测生物分子之间的相互作用一直是生物医学科学的一个主要重点领域，因为这种相互作用支配着生物体的所有过程。检测生物分子之间结合的能力对于新疗法和治疗剂的开发至关重要。这项提议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.