Collaborative Research: Molecular basis for protein sorption in polymer-modified chromatographic media

Lenhoff/Szleifer1263966/1264696合作项目的目标是量化聚合物衍生介质(PDM)的功能特征，以通过实验和计算方法了解蛋白质在基质中的吸附和传输机制。重点放在既允许分配又允许表面吸附的离子交换聚合物上。这项研究围绕三个具体目标展开：1)确定蛋白质在不同结构的PDM上的比较吸附和传输特性。2)测量吸附过程和吸附状态的微观特征，作为识别和分析吸附和传输机制的指导。3)推广和应用分子理论来描述蛋白质在PDM中的吸附平衡和动力学。前两个目标包括产生大量的实验数据，使详细的机理分析成为可能。这两个目标的不同之处在于，第一个目标涵盖在正常工艺条件下对评估功能有直接意义的色谱行为，而第二个目标涉及更细粒度的、主要是微观方法，旨在解决关于可能机制的歧义。第三个目标将使用现实的、最先进的分子建模来模拟蛋白质在聚合物衍生的离子交换介质中的平衡和动力学行为，并为解决机理上的不确定性提供严格的基础。一旦完成，这些研究将允许开发适用于色谱建模的更简单的关系，然后这些方法将被应用于预测模型混合物的分离效果。研究的成功完成将产生结构-功能关系，有助于解释观察到的性能趋势，优化特定分离的材料选择，并设计新型吸附剂。更广泛的影响。该项目的结果有望阐明产品数据管理的作用机制，从而有利于生物技术和制药行业的研究和开发。这项工作可能有助于解决目前困扰某些应用的一些重要问题，如洗脱速度慢。对于固定相制造商来说，对基本作用机制的更清楚理解可以指导新吸附剂的开发；PI与相关公司进行了合作。虽然研究结果将主要通过在同行评议的期刊上发表来传播，但私人投资机构也将向研究界提供更多的研究成果，包括将分子理论应用于色谱过程的软件。多学科和多地点合作将丰富学生研究培训的影响。

Lenhoff / Szleifer1263966 / 1264696The goal of the collaborative project is to quantify the functional characteristics of polymer-derivatized media (PDM) to gain an understanding of the mechanism of protein sorption and transport within the matrix using experimental and computational methods. The focus is on ion exchange polymers that allow for both partitioning and surface sorption. The research is organized around three specific aims:1) Determine comparative sorption and transport characteristics for proteins on PDM with different architectures.2) Measure microscopic characteristics of the sorption process and the sorbed state as a guide to identification and analysis of sorption and transport mechanisms.3) Generalize and apply molecular theories to describe the sorption equilibrium and kinetics of proteins in PDM.The first two aims cover generation of a wide array of experimental data that will make possible a detailed mechanistic analysis of the phenomena involved. These two aims differ in that the first covers chromatographic behavior that is directly meaningful in assessing function under normal process conditions, while the second involves more fine-grained, primarily microscopic methods that are intended to resolve ambiguities regarding possible mechanisms. The third aim will employ realistic, state-of-the art molecular modeling to simulate equilibrium and kinetic behavior of proteins in polymer-derivatized ion-exchange media and provide a rigorous basis for resolving mechanistic uncertainties. Once completed, these studies will allow the development of simpler relationships suitable for routine use in chromatographic modeling, and these methods will then be applied to predicting the efficacy of separation of a model mixture. Successful completion of the investigation will yield structure-function relations that can facilitate explanation of observed performance trends, optimization of materials selection for specific separations, and design of novel adsorbents. Broader impacts. The results of the project are expected to elucidate the mechanisms of actions of PDM and hence benefit research and development in the biotechnology and pharmaceutical industries. The work may help resolve important issues such as slow elution that currently bedevil some applications. For stationary-phase manufacturers, a clearer understanding of the underlying mechanisms of action can guide development of new adsorbents; the PIs have collaborations with relevant companies. While the findings of the research will be disseminated mainly by publication in peer-reviewed journals, the PIs will also make the larger output from the research available to the research community, including software to apply the molecular theory to chromatographic processes. The impact in student training in research will be enriched by the multidisciplinary and multisite collaboration.