title: Spermine Sepharose as a clustered-charge anion exchange adsorbent.
abstract: 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 α-lactalbumin, and RNA from baker's yeast, were adsorbed on a spermine Sepharose adsorbent. This adsorbent exhibited enhanced α-lactalbumin binding capacity (Qmax>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 (Qmax/KD 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.
authors.affiliation: Department of Biology & Biochemistry, University of Houston, Houston, TX 77204-5001, USA.
authors.forename: Sagar
authors.initials: S
authors.lastname: Dhamane
authors.affiliation: Department of Biology & Biochemistry, University of Houston, Houston, TX 77204-5001, USA; Centro de Biotecnología FEMSA, Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico.
authors.forename: Federico
authors.initials: F
authors.lastname: Ruiz-Ruiz
authors.affiliation: Department of Chemical & Biomolecular Engineering, University of Houston, Houston, TX 77204-4004, USA.
authors.forename: Wen-hsiang
authors.initials: WH
authors.lastname: Chen
authors.affiliation: Department of Chemical & Biomolecular Engineering, University of Houston, Houston, TX 77204-4004, USA.
authors.forename: Katerina
authors.initials: K
authors.lastname: Kourentzi
authors.affiliation: Centro de Biotecnología FEMSA, Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico.
authors.forename: Jorge
authors.initials: J
authors.lastname: Benavides
authors.affiliation: Centro de Biotecnología FEMSA, Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico.
authors.forename: Marco
authors.initials: M
authors.lastname: Rito-Palomares
authors.affiliation: Department of Biology & Biochemistry, University of Houston, Houston, TX 77204-5001, USA; Centro de Biotecnología FEMSA, Departamento de Biotecnología e Ingeniería de Alimentos, Tecnológico de Monterrey, Campus Monterrey, Ave. Eugenio Garza Sada 2501 Sur, Monterrey, NL 64849, Mexico; Department of Chemical & Biomolecular Engineering, University of Houston, Houston, TX 77204-4004, USA. Electronic address: willson@uh.edu.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Willson
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D007768:Lactalbumin / Q000302:isolation & purification; D008024:Ligands; D012331:RNA, Fungal / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D012685:Sepharose / Q000737:chemistry*; D013096:Spermine / Q000737:chemistry*
chemical_list: D008024:Ligands; D012331:RNA, Fungal; D013096:Spermine; D012685:Sepharose; D007768:Lactalbumin
keywords: Adsorption; Clustered charge; Ion-exchange chromatography; Ligand density; Spermine