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Bioassays and Molecular Characterization by Microchip CE

通过 Microchip CE 进行生物测定和分子表征

基本信息

批准号：
7163832
负责人：
PATRICK A LIMBACH
金额：
$ 32.1万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-01-01 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7163832
关键词：
Acids Affinity Biological Biological Assay Capillary Electrophoresis Categories Chemicals Communities Compatible Complex Consumption Coupled Cycloparaffins Detection Development Disease Disease Progression Dissociation Effectiveness Electrospray Ionization Equilibrium Fluorescence Goals Hour Immunoassay Intervention Investigation Lasers Ligand Binding Mass Spectrum Analysis Medical Medical Research Methodology Methods Methylmethacrylate Modeling Molecular Monitor Nucleic Acids Octanols Operative Surgical Procedures Optics Performance Pharmacologic Substance Plastics Preclinical Drug Evaluation Preparation Principal Investigator Process Production Property Proteins Range Research Research Project Grants Resolution Sampling Scheme Speed Standards of Weights and Measures System Techniques Technology Therapeutic Time Water aptamer base charge coupled device camera copolymer cost design detector improved microchip physical property polycarbonate polypeptide programs small molecule

项目摘要

DESCRIPTION (provided by applicant): The goal of this research is to develop plastic microchip analytical technologies that yield high-throughput approaches for the trace determination and/or physical properties of biological and small molecule therapeutics that are increasingly important to pharmaceutical and medical research. In support of this long-term goal are two specific aims. The first aim is to develop appropriate assays from three categories that would benefit significantly from the advantages of the microchip platform. The research focus will include efforts to significantly improve the determination of trace amounts of biological compounds with respect to speed and selectivity as compared with the standard immunoassay methodology. Aptamer-based assays of proteins will be used as illustrative chemical systems, and capillary electrophoresis on a multilane plastic chip with detection by laser induced fluorescence (LIF) will be the analytical technique. Analysis time will be shortened from hours to minutes by rapid, high-resolution separation of protein-photoaptamer complexes without the need for prior sample preparation. Additionally, a relatively high-throughput non-equilibrium separation-based approach that allows the accurate determination of Kd and IC-50 values from a wide range of ligand-binding systems will be developed. Non-equilibrium affinity capillary electrophoresis performed on a multilane plastic microchip with LIF, indirect LIF or flow-assisted mass spectrometry detection will be used with several model ligand-binding systems. As the final illustration, a universal, low sample consumption, high throughput and low cost separation-based platform to determine two of the key physiochemical properties of small molecules (MW 200-1,000), pKa and log Pow will be developed. Multilane plastic microchips coupled with either indirect laser-induced fluorescence or mass spectrometric detection can provide a rapid means of assessing these key parameters. The second aim will be to develop appropriate plastic microchip platforms from which these assays can be performed. In particular, these platforms must be compatible with the assays developed in the first Specific Aim. Efforts will be directed at developing multilane plastic chips having appropriate properties for use with LIF or indirect LIF as well as the appropriate laser system for LIF-associated methods. Efforts will also focus on appropriate chip designs and plastic substrates for multi-analyte analyses with mass spectrometric detection methods. Upon completion, appropriate microchip-sets will be created for assays of biological and small molecule therapeutics.

描述（由申请人提供）：

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Comparing polyelectrolyte multilayer-coated PMMA microfluidic devices and glass microchips for electrophoretic separations.

DOI：
10.1002/elps.200900403
发表时间：
2009-12
期刊：
ELECTROPHORESIS
影响因子：
2.9
作者：
Currie, Christa A.;Shim, Joon Sub;Lee, Se Hwan;Ahn, Chong;Limbach, Patrick A.;Halsall, H. Brian;Heineman, William R.
通讯作者：
Heineman, William R.

Parallel separations using capillary electrophoresis on a multilane microchip with multiplexed laser-induced fluorescence detection.

DOI：
10.1002/elps.201000030
发表时间：
2010-08
期刊：
ELECTROPHORESIS
影响因子：
2.9
作者：
Nikcevic, Irena;Piruska, Aigars;Wehmeyer, Kenneth R.;Seliskar, Carl J.;Limbach, Patrick A.;Heineman, William R.
通讯作者：
Heineman, William R.

Unlimited-volume electrokinetic stacking injection in sweeping capillary electrophoresis using a cationic surfactant.

使用阳离子表面活性剂在扫频毛细管电泳中进行无限体积电动堆积注射。