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

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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.