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Updated: Sep 3, 2025

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Electrospun Hydrophobic Interaction Chromatography (HIC) Membranes for Protein Purification
Shu-Ting Chen1, Sumith Ranil Wickramasinghe1, Xianghong Qian2
1Department of Chemical Engineering, University of Arkansas, Fayetteville, AR 72701, USA.
Abstract:
Responsive membranes for hydrophobic interaction chromatography have been fabricated by functionalizing poly(N-vinylcaprolactam) (PVCL) ligands on the substrate of electrospun regenerated cellulose nanofibers. Both static and dynamic binding capacities and product recovery were investigated using bovine serum albumin (BSA) and Immunoglobulin G (IgG) as model proteins. The effects of ligand chain length and chain density on static binding capacity were also studied. A static binding capacity of ~25 mg/mL of membrane volume (MV) can be achieved in optimal ligand grafting conditions. For dynamic binding studies, protein binding capacity increased with protein concentration from 0.1 to 1.0 g/L. Dynamic binding capacity increased from ~8 mg/mL MV at 0.1 g/L BSA to over 30 mg/mL at 1.0 g/L BSA. However, BSA recovery decreased as protein concentration increased from ~98% at 0.1 g/L BSA to 51% at 1 g/L BSA loading concentration. There is a clear trade-off between binding capacity and recovery rate. The electrospun substrate with thicker fibers and more open pore structures is superior to thinner fibrous membrane substrates.
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