Fluorescence correlation spectroscopy study of protein transport and dynamic interactions with clustered-charge peptide adsorbents.
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.
Department of Chemistry, Rice University, Houston, TX 77251, USA.
Charlisa R
CR
Daniels
Lydia
L
Kisley
Hannah
H
Kim
Wen-Hsiang
WH
Chen
Mohan-Vivekanandan
MV
Poongavanam
Carmen
C
Reznik
Katerina
K
Kourentzi
Richard C
RC
Willson
Christy F
CF
Landes
D000327:Adsorption; D002852:Chromatography, Ion Exchange; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D010455:Peptides; D011506:Proteins