TY - JOUR
T1 - Fluorescence correlation spectroscopy study of protein transport and dynamic interactions with clustered-charge peptide adsorbents
AU - Daniels, Charlisa R.
AU - Kisley, Lydia
AU - Kim, Hannah
AU - Chen, Wen Hsiang
AU - Poongavanam, Mohan Vivekanandan
AU - Reznik, Carmen
AU - Kourentzi, Katerina
AU - Willson, Richard C.
AU - Landes, Christy F.
N1 - Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2012/8
Y1 - 2012/8
N2 - 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.
AB - 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.
KW - anion-exchange matrix
KW - clustered-charge ion-exchange adsorbent
KW - diffusion
KW - ion-exchange chromatography
KW - protein adsorption
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U2 - 10.1002/jmr.2206
DO - 10.1002/jmr.2206
M3 - Article
C2 - 22811068
AN - SCOPUS:84864200059
SN - 0952-3499
VL - 25
SP - 435
EP - 442
JO - Journal of Molecular Recognition
JF - Journal of Molecular Recognition
IS - 8
ER -