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Effects of molecule size and resin structure on protein adsorption on multimodal anion exchange chromatography media.

Authors
  • Roberts, Joey A1
  • Kimerer, Lucas1
  • Carta, Giorgio2
  • 1 Department of Chemical Engineering, University of Virginia, Charlottesville, VA, USA.
  • 2 Department of Chemical Engineering, University of Virginia, Charlottesville, VA, USA. Electronic address: [email protected]
Type
Published Article
Journal
Journal of chromatography. A
Publication Date
Sep 27, 2020
Volume
1628
Pages
461444–461444
Identifiers
DOI: 10.1016/j.chroma.2020.461444
PMID: 32822983
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

The effect of bead and ligand structure on protein adsorption was investigated for multimodal anion exchangers combining a quaternary ammonium ion group with hydrophobic moieties: Nuvia aPrime 1 and aPrime 2, based on a 54 μm diameter polymeric bead, and Capto Adhere ImpRes and Capto Adhere, based on agarose beads 51 and 78 μm diameter, respectively. Bovine serum albumin (BSA) monomer, BSA dimer, and thyroglobulin (Tg) were used as model proteins. Based on TEM imaging and iSEC, the Nuvia resins have a microgranular structure and large pores (110 nm radius), while the Capto resins have a fibrous structure and smaller pores (32-36 nm radius). Comparable binding capacities (80-110 mg/mL), decreasing as salt is added, are observed for all three proteins on the Nuvia resins. Higher capacities (110-130 mg/mL), also decreasing as salt is added, are observed for BSA monomer and dimer on the Capto resins. However, the Tg binding capacity is very low in this case and increases as salt is added. Confocal laser scanning microscopy show that the kinetics are controlled by pore diffusion for all four resins, but with diffusivities that decrease as the protein size increases especially for the Capto resins. For Tg at low salt, binding is restricted to a thin shell close to the bead surface for both Capto resins. The ratio of effective and free diffusivity is about 0.30, 0.18, and 0.08 for BSA monomer, BSA dimer, and Tg, respectively, on the Nuvia resin. These values decrease to about 0.11, 0.04, and 0.01, respectively, for the Capto resins as a result of diffusional hindrance. Dynamic binding capacities are consistent with the equilibrium and rate behaviors. Copyright © 2020. Published by Elsevier B.V.

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