Microcantilever Biosensors Based on Ligand Gated Ion Channel Receptors

基于配体门控离子通道受体的微悬臂生物传感器

• 批准号: 7797015
• 负责人: Marvin Kenneth Schulte
• 金额: $ 1.44万
• 依托单位: UNIVERSITY OF ALASKA FAIRBANKS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7797015
• 关键词: Acetylcholine; Binding; Binding Proteins; Biological Assay; Biosensor; Brain; Chemistry; Coupled; Data; Detection; Diagnostic; Drug Evaluation; Drug Receptors; Environmental Monitoring; Family; Gated Ion Channel; Goals; HTR3A gene; Histamine; Human; Ion Channel Protein; Laboratories; Ligands; Modeling; Modification; Molecular Models; Mutate; Neurons; Nicotinic Receptors; Noise; Performance; Peripheral Nervous System; Pharmacology; Preclinical Drug Evaluation; Protein Family; Proteins; Protocols documentation; Reproducibility; Research; Screening procedure; Sensitivity and Specificity; Series; Serotonin; Signal Transduction; Site-Directed Mutagenesis; Specificity; Structure; Surface; System; Technology; Testing; Therapeutic Agents; Time; Vision; Work; base; cantilever; density; design; drug candidate; drug development; drug discovery; gamma-Aminobutyric Acid; improved; knowledge base; member; molecular modeling; novel; protocol development; receptor; response; sensor; serotonin receptor; serotonin-binding protein; simulation

DESCRIPTION (provided by applicant): The primary goal of this project is to develop a sensor for use in drug development that mimics human brain receptors. This sensor will be capable of evaluating the binding of large numbers of drug candidates in a short period of time; thus greatly enhancing the rate at which potential therapeutic agents can be identified. The primary goal of this project is to develop improved microcantilever-based biosensors using receptor proteins based on a recently discovered acetylcholine binding protein (AChBP). The AChBP is a soluble protein that displays structure and pharmacology that is strikingly similar to the nicotinic acetylcholine receptor (nAChR) present in the central and peripheral nervous systems. Since the nAChR is a member of a large super-family of ligand gated ion channels (LGICs), this project will ultimately develop sensors that will be useful in high throughput drug screening and drug discovery for this class of neuronal receptors. Specifically, we aim to: 1) Develop a microcantilever biosensor utilizing the AChBP and its derivatives as biological sensor proteins. 2) Investigate fundamental surface conjugation chemistries and establish modification protocols for microcantilever sensing. The project draws heavily on our combined expertise in LGIC structure and function (Dr. Schulte) and microcantilever design (Dr. Ji). Aim 1 will evaluate the AChBP itself as a potentially useful biosensor molecule. We will initially demonstrate the use of this sensor in high-throughput drug screening by exploring its interaction with nAChR, GABAR, glycineR and 5-HT3R ligands. Since the AChBP can be mutated to alter its ligand specificity, derivatives obtained via site directed mutagenesis and/or chimeric construction will also be developed. Since the AChBP is commonly used as a template for modeling of ligand gated ion channel receptors, an additional benefit of this work will be an expanded knowledge of the basis of ligand specificity at these important receptors. Aim 2 will utilize the AChBP and other receptor proteins to improve sensor response through improved conjugation chemistries in combination with improved microcantilever array technology developed in Dr. Ji's laboratory

描述（由申请人提供）：该项目的主要目标是开发一种用于模拟人脑受体的药物开发中的传感器。该传感器将能够在短时间内评估大量候选药物的结合；因此，可以大大提高潜在的治疗剂的速度。该项目的主要目的是使用基于最近发现的乙酰胆碱结合蛋白（ACHBP）的受体蛋白（ACHBP）开发改进的基于微抗体的生物传感器。 ACHBP是一种可溶性蛋白，显示出与中央和周围神经系统中存在的烟碱乙酰胆碱受体（NACHR）非常相似的结构和药理学。由于NACHR是配体门控离子通道（LGICS）的大型超级家庭的成员，因此该项目最终将开发该类别的神经元受体的高通量药物筛查和药物发现的传感器。具体而言，我们的目的是：1）使用ACHBP及其衍生物作为生物传感器蛋白的微型管理器生物传感器。 2）研究基本的表面共轭化学并建立用于微抗体传感的修改方案。该项目在LGIC结构和功能（Schulte Dr.）和MicroCantilever Design（JI博士）方面的综合专业知识很大程度上借鉴了。 AIM 1将评估ACHBP本身作为潜在有用的生物传感器分子。我们最初将通过探索其与NACHR，GABAR，甘同能和5-HT3R配体的相互作用来证明该传感器在高通量药物筛查中的使用。由于可以将ACHBP突变以改变其配体特异性，因此还将开发通过位置的诱变和/或嵌合构建获得的衍生物。由于ACHBP通常用作建模配体门控离子通道受体的模板，因此这项工作的另一个好处将是对这些重要受体配体特异性基础的扩展知识。 AIM 2将利用ACHBP和其他受体蛋白来通过改进的共轭化学结合结合改进的Microcantilever阵列技术来改善传感器的响应。