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: 7286095
• 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 2009-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7286095
• 关键词: Algorithms; Altretamine; Antibodies; Antigenic Specificity; Antigens; Architecture; Base Pairing; Binding; Biological; Biophysics; Biosensing Techniques; Buffers; Class; Classification; Computer software; Condition; DNA; Data; Detection; Development; Devices; Dissociation; Failure; Feedback; Fingerprint; Haptens; Hemolysin; Immunology; Informatics; Kinetics; Knowledge; Lead; Length; Machine Learning; Measurement; Measures; Methods; Motion; Noise; Pattern Recognition; Physiological; Play; Pliability; Preparation; Principal Investigator; Probability; Process; Proteins; Protocols documentation; Range; Rate; Research; Research Personnel; Resistance; Resolution; Role; Sampling; Screening procedure; Semiconductors; Signal Transduction; Staging; Statistical Methods; Testing; Time; Time Study; Toxin; Variant; antibody engineering; antigen antibody binding; antigen binding; antimicrobial peptide; base; cheminformatics; computerized data processing; design; detector; expectation; experimental analysis; markov model; molecular dynamics; nanopore; programs; protein structure function; prototype; single molecule; stem; tool; vector; web interface

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.

产品说明： 基于纳米孔的单分子检测最近已经成为单分子生物物理学中的新工具。有证据表明，单抗体可以观察到与纳米孔检测器，这提出了广泛的免疫学研究的可能性。待测试的假设是基于纳米孔的检测可用于研究抗体-抗原相互作用的单分子动力学，并分析抗体与抗原结合后发生的构象变化。本申请旨在通过改进检测设备和用于从通道电流信号提取信息的软件来开发基于纳米孔的方法的实用性。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基于纳米孔/抗体的检测。 2.实施机器学习软件，用于自动化纳米孔/抗体信号分析和实验反馈。 3.使用充分表征的基因工程抗体来测试纳米孔装置分析抗体分子运动的效用。 这些研究将扩大纳米孔装置的实用性，以研究单分子蛋白质相互作用。 所获得的信息将导致更好地理解与抗体结合抗原和随后启动效应子功能相关的分子动力学。由于大多数生物纳米孔变体来自成孔毒素，纳米孔装置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用作高灵敏度的免疫传感器。

项目成果

Duration learning for analysis of nanopore ionic current blockades.

用于分析纳米离子电流阻滞的持续时间学习。

• DOI: 10.1186/1471-2105-8-s7-s14
• 发表时间: 2007-11-01
• 期刊: BMC BIOINFORMATICS
• 影响因子: 3
• 作者: Churbanov, Alexander;Baribault, Carl;Winters-Hilt, Stephen
• 通讯作者: Winters-Hilt, Stephen

A novel, fast, HMM-with-Duration implementation - for application with a new, pattern recognition informed, nanopore detector.

• DOI: 10.1186/1471-2105-8-s7-s19
• 发表时间: 2007-11-01
• 期刊: BMC bioinformatics
• 影响因子: 3
• 作者: Winters-Hilt S;Baribault C
• 通讯作者: Baribault C

Nanopore-based kinetics analysis of individual antibody-channel and antibody-antigen interactions.

基于纳米孔的单个抗体通道和抗体-抗原相互作用的动力学分析。

• DOI: 10.1186/1471-2105-8-s7-s20
• 发表时间: 2007
• 期刊: BMC bioinformatics
• 影响因子: 3
• 作者: Winters-Hilt,Stephen;Morales,Eric;Amin,Iftekhar;Stoyanov,Alexander
• 通讯作者: Stoyanov,Alexander

Pattern recognition-informed feedback for nanopore detector cheminformatics.

• DOI: 10.1007/978-1-4419-5913-3_12
• 发表时间: 2010
• 期刊: Advances in experimental medicine and biology
• 作者: A. M. Eren;Iftekhar Amin;Amanda Alba;Eric Morales;A. Stoyanov;S. Winters-Hilt;S. Winters-Hilt

The alpha-hemolysin nanopore transduction detector - single-molecule binding studies and immunological screening of antibodies and aptamers.

α-溶血素纳米孔转导检测器 - 单分子结合研究以及抗体和适体的免疫筛选。

• DOI: 10.1186/1471-2105-8-s7-s9
• 发表时间: 2007
• 期刊: BMC bioinformatics
• 影响因子: 3
• 作者: Winters-Hilt,Stephen