Unified superresolution experiments and stochastic theory provide mechanistic insight into protein ion-exchange adsorptive separations.

ABSTRACT: Chromatographic protein separations, immunoassays, and biosensing all typically involve the adsorption of proteins to surfaces decorated with charged, hydrophobic, or affinity ligands. Despite increasingly widespread use throughout the pharmaceutical industry, mechanistic detail about the interactions of proteins with individual chromatographic adsorbent sites is available only via inference from ensemble measurements such as binding isotherms, calorimetry, and chromatography. In this work, we present the direct superresolution mapping and kinetic characterization of functional sites on ion-exchange ligands based on agarose, a support matrix routinely used in protein chromatography. By quantifying the interactions of single proteins with individual charged ligands, we demonstrate that clus

SUBMITTER: Kisley L 

PROVIDER: S-EPMC3926075 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature