Fluorescence correlation spectroscopy study of protein transport and dynamic interactions with clustered-charge peptide adsorbents

Charlisa R. Daniels, Lydia Kisley, Hannah Kim, Wen Hsiang Chen, Mohan Vivekanandan Poongavanam, Carmen Reznik, Katerina Kourentzi, Richard C. Willson, Christy F. Landes

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Ion-exchange chromatography relies on electrostatic interactions between the adsorbent and the adsorbate and is used extensively in protein purification. Conventional ion-exchange chromatography uses ligands that are singly charged and randomly dispersed over the adsorbent, creating a heterogeneous distribution of potential adsorption sites. Clustered-charge ion exchangers exhibit higher affinity, capacity, and selectivity than their dispersed-charge counterparts of the same total charge density. In the present work, we monitored the transport behavior of an anionic protein near clustered-charge adsorbent surfaces using fluorescence correlation spectroscopy. We can resolve protein-free diffusion, hindered diffusion, and association with bare glass, agarose-coated, and agarose-clustered peptide surfaces, demonstrating that this method can be used to understand and ultimately optimize clustered-charge adsorbent and other surface interactions at the molecular scale.

Original languageEnglish (US)
Pages (from-to)435-442
Number of pages8
JournalJournal of Molecular Recognition
Volume25
Issue number8
DOIs
StatePublished - Aug 2012

Keywords

  • anion-exchange matrix
  • clustered-charge ion-exchange adsorbent
  • diffusion
  • ion-exchange chromatography
  • protein adsorption

ASJC Scopus subject areas

  • Molecular Biology
  • Structural Biology

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