ArticleProceedings of the National Academy of Sciences of the United States of America2014
Unified superresolution experiments and stochastic theory provide mechanistic insight into protein ion-exchange adsorptive separations.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Role of viral protein ratio in the structure and separation of empty and full adeno-associated virus capsids: A molecular dynamics study.Molecular therapy. Advances · 2026Article
- Mechanistic Insights into AAV Capsid-Stationary Phase Interactions Governing Native Stability and Chromatographic Separation Using AAV8 as a Model System.Pharmaceutics · 2026Article
- Super-resolution imaging reveals resistance to mass transfer in functionalized stationary phases.Science advances · 2025Article
- A mechanistic examination of salting out in protein-polymer membrane interactions.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Ultrafast Transient Absorption Spectra of Photoexcited YOYO-1 molecules call for additional investigations of their fluorescence quenching mechanism.Journal of photochemistry and photobiology. A, Chemistry · 2018Article
- Photobleaching of YOYO-1 in super-resolution single DNA fluorescence imaging.Beilstein journal of nanotechnology · 2017Article
- Generalized recovery algorithm for 3D super-resolution microscopy using rotating point spread functions.Scientific reports · 2016Article
- Article
- Characterization of Porous Materials by Fluorescence Correlation Spectroscopy Super-resolution Optical Fluctuation Imaging.ACS nano · 2015Article
- Error-based extraction of states and energy landscapes from experimental single-molecule time-series.Scientific reports · 2015Article
- Molecular approaches to chromatography using single molecule spectroscopy.Analytical chemistry · 2015Review
- Fast Step Transition and State Identification (STaSI) for Discrete Single-Molecule Data Analysis.The journal of physical chemistry letters · 2014Article
- Charge-dependent transport switching of single molecular ions in a weak polyelectrolyte multilayer.Langmuir : the ACS journal of surfaces and colloids · 2014Article
- High ionic strength narrows the population of sites participating in protein ion-exchange adsorption: a single-molecule study.Journal of chromatography. A · 2014Article
- Article
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Authors and funding
10 authors.
Funding
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 clusters of charges are necessary to create detectable adsorption sites and that even chemically identical ligands create adsorption sites of varying kinetic properties that depend on steric availability at the interface. Additionally, we relate experimental results to the stochastic theory of chromatography. Simulated elution profiles calculated from the molecular-scale data suggest that, if it were possible to engineer uniform optimal interactions into ion-exchange systems, separation efficiencies could be improved by as much as a factor of five by deliberately exploiting clustered interactions that currently dominate the ion-exchange process only accidentally.
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