Nanopore study of single antibody-antigen interactions

单一抗体-抗原相互作用的纳米孔研究

• 批准号: 6959048
• 负责人: STEPHEN WINTERS-HILT
• 金额: $ 16.2万
• 依托单位: CHILDREN'S HOSPITAL (NEW ORLEANS)
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2008-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6959048
• 关键词: antigen antibody reaction; bioinformatics; biomedical automation; computer program /software; computer system design /evaluation; intermolecular interaction; molecular dynamics; nanotechnology; pore forming protein; protein quantitation /detection

DESCRIPTION: Nanopore-based single-molecule detection has recently become established as a new tool in single molecule biophysics. Evidence is presented that single antibodies can be observed with a nanopore detector, which presents a wide range of possibilities for immunological research. The hypothesis to be tested is that nanopore-based detection can be used to study single molecule dynamics of antibody-antigen interaction and analyze conformational changes that occur in antibody upon binding to antigen. This application aims to develop the utility of the nanopore-based approach through improvements in both the detection device and the software used to extract information from the channel current signal. At the same time, these studies will allow the Candidate to gain expertise in immunology and the biophysical study of protein structure and function. To study the single molecule dynamics of antibody-antigen interaction, the following three specific aims are proposed: 1. Extend nanopore based detection to nanopore/antibody based detection. 2. Implement machine learning software for automated nanopore/antibody signal analysis and experimental feedback. 3. Use well-characterized, genetically engineered, antibodies to test the utility of the nanopore device to analyze motion in the antibody molecule. These studies will expand the utility of nanopore devices to study single molecule protein interactions. Information gained will lead to a better understanding of the molecular dynamics associated with antigen binding by antibody and the subsequent initiation of effector functions. Since most biological nanopore variants derive from pore-forming toxins, nanopore device enhancements eventually may lead to new methods for antibody and antimicrobial-peptide immunological screening. Antibody-based nanopore devices may also serve as highly sensitive immunosensors.

说明： 近年来，基于纳米孔的单分子检测已经成为单分子生物物理学中的一种新工具。有证据表明，用纳米孔检测器可以观察到单一抗体，这为免疫学研究提供了广泛的可能性。需要检验的假设是，基于纳米孔的检测可以用于研究抗体-抗原相互作用的单分子动力学，并分析抗体与抗原结合时发生的构象变化。这项申请旨在通过改进检测设备和用于从通道电流信号中提取信息的软件来开发基于纳米孔的方法的实用性。与此同时，这些研究将使候选人获得免疫学和蛋白质结构和功能的生物物理研究方面的专业知识。为了研究抗体-抗原相互作用的单分子动力学，提出了以下三个具体目标： 1.将基于纳米孔的检测扩展到纳米孔/抗体的检测。 2.实现自动纳米孔/抗体信号分析和实验反馈的机器学习软件。 3.使用特征良好的基因工程抗体来测试纳米孔设备分析抗体分子运动的效用。 这些研究将扩大纳米孔设备研究单分子蛋白质相互作用的用途。 所获得的信息将有助于更好地理解与抗体结合抗原和随后启动效应器功能相关的分子动力学。由于大多数生物纳米孔变体来自于致孔毒素，纳米孔装置的增强最终可能导致抗体和抗菌肽免疫筛选的新方法。基于抗体的纳米孔装置也可以作为高度敏感的免疫传感器。