Characterization of a macroporous epoxy-polymer based resin for the ion-exchange chromatography of therapeutic proteins

J Chromatogr A. 2021 Oct 25:1656:462503. doi: 10.1016/j.chroma.2021.462503. Epub 2021 Sep 1.

Abstract

This study investigated the adsorption capacity and mass transfer properties of a novel macroporous epoxy-polymer-based anion-exchanger, MPR Q, for the efficient separation of therapeutic proteins. MPR Q resin was prepared by phase separation based on spinodal decomposition followed by dextran grafting and ligand conjugation. Under static conditions, MPR Q exhibited a binding capacity of 49.8 mg-IgG/cm3-resin at pH 10, whereas the fastest adsorption was observed among the anion-exchanger resins tested. Inverse size-exclusion chromatography (iSEC) experiments revealed that the apparent pore diameter of MPR Q was approximately 90 nm, which was sufficiently large for the penetration of human IgG and bovine IgM. Moreover, the reduced height equivalent to a theoretical plate, h, of human IgG, determined using the linear gradient elution method was 65.8 and was not significantly changed in the range of linear velocities from 20.37 to 50.93 cm/min. The dynamic binding capacity at 10% breakthrough of MPR Q, determined by frontal analysis, exhibited a capacity of 43.8 mg/cm3 at 5.09 cm/min and 58% of DBC10% was maintained even though the linear velocity was increased to 50.93 cm/min. Furthermore, a resolution for separation of IgG and BSA by MPR Q was 1.06 at 5.09 cm/min, while it was higher than that for the conventional resin at all linear velocities from 5.09 cm/min to 50.93 cm/min. Thus, it was suggested that the MPR Q developed in this study is a promising resin that can efficiently separate large biomacromolecules such as human IgG at higher velocities.

Keywords: Adsorption kinetics; Dynamic binding capacity; Epoxy-polymer based resin; Height equivalent to a theoretical plate; Ion-exchange chromatography; Mass transfer.

MeSH terms

  • Adsorption
  • Animals
  • Cattle
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Humans
  • Polymers*
  • Serum Albumin, Bovine*

Substances

  • Polymers
  • Serum Albumin, Bovine