Facile preparation of salt-tolerant anion-exchange membrane adsorber using hydrophobic membrane as substrate

Jinxin Fan; Jianquan Luo; Xiangrong Chen; Yinhua Wan

doi:10.1016/j.chroma.2017.02.016

Facile preparation of salt-tolerant anion-exchange membrane adsorber using hydrophobic membrane as substrate

J Chromatogr A. 2017 Mar 24:1490:54-62. doi: 10.1016/j.chroma.2017.02.016. Epub 2017 Feb 14.

Authors

Jinxin Fan¹, Jianquan Luo², Xiangrong Chen³, Yinhua Wan⁴

Affiliations

¹ State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, University of the Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
² State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, University of the Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100190, PR China. Electronic address: jqluo@ipe.ac.cn.
³ State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, University of the Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100190, PR China.
⁴ State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, University of the Chinese Academy of Sciences, Chinese Academy of Sciences, Beijing 100190, PR China. Electronic address: yhwan@ipe.ac.cn.

PMID: 28215404
DOI: 10.1016/j.chroma.2017.02.016

Abstract

In this study, a polyvinylidene fluoride (PVDF) hydrophobic membrane with high mechanical property was used as substrate to prepare salt-tolerant anion-exchange (STAE) membrane adsorber. Effective hydrophilization and functionalization of PVDF membrane was realized via polydopamine (PDA) deposition, thus overcoming the drawbacks of hydrophobic substrates including poor water permeability, inert property as well as severe non-specific adsorption. The following polyallylamine (PAH) coupling was carried out at pH 10.0, where unprotonated amine groups on PAH chains were more prone to couple with PDA. This membrane adsorber could remain 75% of protein binding capacity when NaCl concentration increased from 0 to 150mM, while its protein binding capacity was independent of flow rate from 10 to 100 membrane volume (MV)/min due to its high mechanical strength (tensile strength: 43.58±2.30MPa). With 200mM NaCl addition at pH 7.5, high purity (above 99%) and high recovery (almost 100%) of Immunoglobulin G (IgG) were obtained when using the STAE membrane adsorber to separate IgG/human serum albumin (HSA) mixture, being similar to that without NaCl at pH 6.0 (both under the flow rate of 10-100MV/min). Finally, the reliable reusability was confirmed by five reuse cycles of protein binding and elution operations. In comparison with commercial membrane adsorber, the new membrane adsorber exhibited a better mechanical property, higher IgG polishing efficiency and reusability, while the protein binding capacity was lower due to less NH₂ density on the membrane. The outcome of this work not only offers a facile and effective approach to prepare membrane adsorbers based on hydrophobic membranes, but also demonstrates great potential of this new designed STAE membrane adsorbers for efficient monoclonal antibody (mAb) polishing.

Keywords: Membrane adsorber; Membrane chromatography; Monoclonal antibody polishing; Mussel-inspired; Salt-tolerant anion-exchange.

MeSH terms

Antibodies, Monoclonal / chemistry
Chromatography, Ion Exchange / methods*
Immunoglobulin G / chemistry
Membranes, Artificial*
Polyvinyls*
Protein Binding

Substances

Antibodies, Monoclonal
Immunoglobulin G
Membranes, Artificial
Polyvinyls
polyvinylidene fluoride