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

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.

Description

Keywords

ion?exchange chromatography, protein adsorption, diffusion, anion?exchange matrix, clustered?charge ion?exchange adsorbent

Citation

Copyright 2012 Journal of Molecular Recognition. This is a post-print version of a published paper that is aviialable at: https://onlinelibrary.wiley.com/doi/full/10.1002/jmr.2206. Recommended citation: Daniels, Charlisa R., Lydia Kisley, Hannah Kim, Wen?Hsiang Chen, Mohan?Vivekanandan Poongavanam, Carmen Reznik, Katerina Kourentzi, Richard C. Willson, and Christy F. Landes. "Fluorescence correlation spectroscopy study of protein transport and dynamic interactions with clustered?charge peptide adsorbents." Journal of Molecular Recognition 25, no. 8 (2012): 435-442. DOI: 10.1002/jmr.2206. This item has been deposited in acordance with the publisher copyright and licensing terms and with the author's permission.