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Preparative Chromatography of Biological Macromolecules

生物大分子的制备色谱

基本信息

批准号：
7194288
负责人：
ABRAHAM M LENHOFF
金额：
$ 21.48万
依托单位：
UNIVERSITY OF DELAWARE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-03-01 至 2009-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7194288
关键词：
3-Dimensional Address Adsorption Area Arts Atomic Force Microscopy Behavior Benchmarking Biological Biological Response Modifier Therapy Biotechnology Characteristics Chromatography Condition Coupled Diffusion Engineering Experimental Models Gene Transduction Agent Goals Image Imaging Techniques Ion-Exchange Chromatography Procedure Laser Scanning Confocal Microscopy Licensing Manufacturer Name Measurement Measures Methods Modeling Modification Monitor Monoclonal Antibodies Outcome Performance Pharmacologic Substance Phase Proteins Range Rate Recovery Research Research Personnel Scanning Sodium Chloride Structure Surface Thermodynamics Vaccines Viral Proteins Virus Virus-like particle Work base bioprocess design electron tomography improved insight macromolecule network models novel programs reconstruction research study simulation size solute theories three dimensional structure uptake

项目摘要

DESCRIPTION (provided by applicant): The proposed research seeks to improve the efficacy of preparative chromatographic separations of large biomacromolecules and assemblies, ranging from large proteins to viruses. Larger proteins such as monoclonal antibodies are increasingly found among licensed biotherapeutics, while viruses and virus-like particles are purified chromatographically as vaccines and as gene therapy vectors. The approaches that will be used to accomplish such improvements are based on developing a better quantitative and qualitative understanding of the mechanisms of adsorption, desorption and transport of large solutes in chromatographic media. The specific aims that will be addressed within this framework of seeking structure-performance relationships will employ chromatographic measurements and modification of stationary-phase surfaces, as well as several imaging methods, viz. electron tomography to determine the 3D structure of pore networks, scanning confocal microscopy to monitor uptake of solutes into stationary phases, and atomic force microscopy to observe the structure of adsorbate layers. The insights obtained from these methods will be coupled with appropriate modeling approaches, including simulations of solute diffusion in pore networks, modeling of uptake into chromatographic media, and colloidal modeling of adsorbate interactions. The experimental and modeling approaches will be used synergistically, and evaluated using several proteins and viruses on a variety of commercial stationary phases that are routinely used in preparative chromatography practice. Successful completion of the proposed research will facilitate efficient and rational approaches to stationary-phase selection for a particular application, and aid in methods for designing novel stationary phases.

描述（由申请人提供）：拟议的研究旨在提高从大型蛋白质到病毒的大型生物大分子和组件的制备层析分离的功效。在获得许可的生物治疗药物中越来越多地发现单克隆抗体等较大的蛋白质，而病毒和病毒样颗粒则作为疫苗和基因治疗载体进行层析纯化。将用于实现这些改进的方法是基于对色谱介质中大溶质的吸附、解吸和运输机制进行更好的定量和定性理解。在寻找结构-性能关系的框架内，将解决的具体目标将采用色谱测量和固定相表面的修改，以及几种成像方法，即电子断层扫描来确定孔隙网络的3D结构，扫描共聚焦显微镜来监测溶质进入固定相的吸收，原子力显微镜来观察吸附层的结构。从这些方法中获得的见解将与适当的建模方法相结合，包括孔隙网络中溶质扩散的模拟，色谱介质摄取的建模，以及吸附质相互作用的胶体模型。实验和建模方法将协同使用，并使用几种蛋白质和病毒在各种商业固定相上进行评估，这些固定相通常用于制备色谱实践。成功完成所提出的研究将有助于为特定应用提供有效和合理的固定相选择方法，并有助于设计新型固定相的方法。