期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1490, 期 -, 页码 54-62出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2017.02.016
关键词
Mussel-inspired; Salt-tolerant anion-exchange; Membrane adsorber; Monoclonal antibody polishing; Membrane chromatography
资金
- National High Technology Research and Development Program of China (863 Program) [2012AA021903, 2014AA021006]
- 100 Talents Program of Youth Innovation Promotion Association Chinese Academy of Sciences [2017069]
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 150 mM, 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 +/- 2.30 MPa). With 200 mM NaCI 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 NaCI at pH 6.0 (both under the flow rate of 10-100 MV/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. (C) 2017 Elsevier B.V. All rights reserved.
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