Journal of Chromatography & Separation Techniques
Open Access
ISSN: 2157-7064
+44 1300 500008
ISSN: 2157-7064
+44 1300 500008
Sena Yaman, Marco A Mata-Gomez, Jose Gonzalez-Valdez, Jesus A Valencia-Gallegos, Marco Rito-Palomares and Canan Tari
Tecnologico de Monterrey, M�©xico
Izmir Institute of Technology, Turkey
Scientific Tracks Abstracts: J Chromatogr Sep Tech
Hydrophobic Interaction Chromatography (HIC) is a powerful technique used to purify proteins. It relies on the interaction between non-polar groups on the hydrophobic patches of the proteins and ligands on the HIC medium. This interaction varies according to the hydrophobicity of biomolecules, leading to the separation of proteins in a complex mixture. In this work, two novel stationary phases for HIC were synthesized by incorporating dendronitic structures, branched molecules. The branched nature of the dendrons allowed the attachment of high amount of hydrophobic ligands to the support material. Depending on the number of branches of the dendron used, ligand densities were calculated as 82.5�±11 and 175.6�±5.7 �¼mol ligand/ml resin for Sorbent 1 and Sorbent 2, respectively. UV-Vis absorption spectra of the modified sorbents exhibited a band at 287 nm corresponding to the aromatic ring present on the dendrons suggesting their incorporation onto the sorbents. FTIR analysis evidenced the aromatic and carbonyl groups suggesting the presecence of dendrons with hidrophobic ligands on the sorbents. Adsorption capacity of the sorbents was evaluated in static and dynamic mode using bovine serum albumin (BSA) under high concentrations of ammonium sulfate (AS). Increasing AS concentration from 1.5-2 M led to significant increases in adsorption capacity. Dynamic adsorption was influenced by flow velocity. This innovative design allowed to increase the ligand density and therefore the adsorption capacity of the sorbents. This technology may permit to reduce the amount of sorbent to be used in a bioprocess and thus use smaller columns resulting in faster chromatoraphic processes.
Sena Yaman has completed her MSc degree in Middle East Technical University and is currently working on her Doctoral studies in Izmir Institute of Technology, Department of Bioengineering.
Email: senaymn@gmail.com