Spermine Sepharose as a clustered-charge anion exchange adsorbent



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Ruiz Ruiz, Fernando and Dhamanea, Sagar and Chen, Wen-Hsiang and Kourentzic, Katerina and Benavides, Jorge and Rito Palomares, Marco and Willson, Richard C. (2014) Spermine Sepharose as a clustered-charge anion exchange adsorbent. Journal of Chromatography A, 1324 . pp. 135-140. ISSN 0021-9673

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Official URL: http://dx.doi.org/10.1016/j.chroma.2013.11.030


We previously showed that the affinity and capacity of ion exchange adsorbents of a given total charge density are improved by immobilization of the charges in pre-ordered clusters, rather than individually in random locations. This previous work used pentalysinamide and pentaargininamide as clustered ligands. This approach allows close control of cluster size, but is uneconomically expensive for some research and most practical applications. In this work, we demonstrate that the inexpensive synthetic analog of the natural polyamine spermine (H2N-CH2-CH2-CH2-NH-CH2-CH2-CH2-CH2-NH-CH2-CH2-CH2 NH2) also can serve as the basis of effective clustered adsorbents. The calcium-depleted form of the protein a lactalbumin, and RNA from baker's yeast, were adsorbed on a spermine Sepharose adsorbent. This adsorbent exhibited enhanced alpha lactalbumin binding capacity (Q(max) >1.6 and 1.3-fold higher than those for Qiagen DEAE and GE DEAE Sepharose adsorbents of much greater charge density) and higher initial binding affinity (Q(max)/K-D 2.4 and 2.1 fold higher, respectively). The new spermine-based matrix exhibited a higher value of the Z parameter, suggesting an increased number of apparent interaction sites between the protein and the resin, and functioned well in column mode. (C) 2013 Elsevier B.V. All rights reserved.

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© 2013 Elsevier B.V. All rights reserved.
This research was funded by the Robert A. Welch foundation under Grant E-1264 and the National Science Foundation (NSF) Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET-1133965). The authors also acknowledge Tecnológico de Monterrey Research chair (Grant CAT161) and CONACyT for fellowship #295368 to FRR.

Uncontrolled Keywords:Thermodynamic Driving Forces; Bovine Alpha-Lactalbumin; Ion-Exchange, Ligand Density, Cytochrome B(5), Molten-Globule, Compact State, Chromatography, Proteins, Adsorption
Subjects:Sciences > Physics
ID Code:25545
Deposited On:28 May 2014 14:41
Last Modified:31 Dec 2020 00:03

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