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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

Postal Subscription Code 80-969

2018 Impact Factor: 2.809

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Front. Chem. Sci. Eng.    2019, Vol. 13 Issue (1) : 70-79    https://doi.org/10.1007/s11705-018-1745-4
RESEARCH ARTICLE
Preparation of copolymer-grafted mixed-mode resins for immunoglobulin G adsorption
Shenggang Chen, Tao Liu, Ruiqi Yang, Dongqiang Lin, Shanjing Yao()
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China
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Abstract

The mixed-mode resins for protein adsorption have been prepared by a novel strategy, copolymer grafting. Specially, the copolymer-grafted resins CG-M-A with two functional groups, 5-amino-benzimidazole (ABI) and methacryloxyethyltrimethyl ammonium chloride (METAC), have been prepared through surface-initiated activator generated by electron transfer for atom transfer radical polymerization of METAC and glycidyl methacrylate (GMA), followed by a ring-open reaction to introduce ABI. The charge and hydrophobicity of CG-M-A resins could be controlled by manipulating the addition of METAC and GMA/ABI. Besides, METAC and ABI provided positive effects together in both protein adsorption and elution: dynamic binding capacity of human Immunoglobulin G (hIgG) onto CG-M-A resin with the highest ligand ratio of METAC to ABI is 46.8 mg·g−1 at pH 9 and the elution recovery of hIgG is 97.0% at pH 5. The separation experiment showed that purity and recovery of monoclonal antibody from cell culture supernatant are 96.0% and 86.5%, respectively, indicating that copolymer-grafted mixed-mode resins could be used for antibody purification.
Keywords atom transfer radical polymerization      copolymer-grafting      mixed-mode resin      protein adsorption     
Corresponding Author(s): Shanjing Yao   
Just Accepted Date: 14 May 2018   Online First Date: 13 September 2018    Issue Date: 25 February 2019
 Cite this article:   
Shenggang Chen,Tao Liu,Ruiqi Yang, et al. Preparation of copolymer-grafted mixed-mode resins for immunoglobulin G adsorption[J]. Front. Chem. Sci. Eng., 2019, 13(1): 70-79.
 URL:  
https://academic.hep.com.cn/fcse/EN/10.1007/s11705-018-1745-4
https://academic.hep.com.cn/fcse/EN/Y2019/V13/I1/70
Fig.1  Illustration of the controlled preparation of copolymer-grafted MMC resins CG-M-A
Resins Amount of monomers Ligand density Reaction efficiency
METAC /mmol?g−1 GMA/ABI
/mmol?g−1		 LDMETAC
/mmol?g−1		 LDABI
/mmol?g−1		 METAC ABI
G-A320 0 1500 0 323±3 - 22.0%
CG-M60-A300 150 1350 56±2 302±4 40.0% 22.2%
CG-M160-A260 375 1125 157±4 259±6 42.7% 23.1%
CG-M330-A230 750 750 328±6 228±5 44.0% 30.7%
Tab.1  The METAC and GMA/ABI amount in the preparation of copolymer-grafted MMC resins, the ligand density of the copolymer-grafted MMC resins and the reaction efficiencies of METAC and ABI
Fig.2  Dextran calibration curves for copolymer-grafted MMC resins CG-M-A and polymer-grafted resin G-A320
Resins rmean/nm e ep eT
G-A320 60.3±1.1 0.42 0.58 0.76
CG-M60-A300 46.6±1.1 0.42 0.57 0.75
CG-M160-A260 38.0±1.1 0.41 0.67 0.81
CG-M330-A230 35.0±2.0 0.35 0.55 0.71
Tab.2  Interstitial, intraparticle and total porosities, and mean pore radii for copolymer-grafted MMC resins at different ligand ratios
Fig.3  Qm values of hIgG on resins G-M-A and CG-M-A with different ligand ratios at different pH
Fig.4  (a) Q10% and (b) Q10%/Q* values of hIgG on resins G-M-A and CG-M-A with different ligand ratios at different pHs
Fig.5  hIgG elution and regeneration profiles on resins CG-M-A and G-A320 using a step elution at pH 5 (20 mmol·L−1) and regeneration at 0.1 mol·L−1 NaOH. Loadings in all cases were 100% of the DBC at 0.65 mL·min−1
Fig.6  Comparison of various samples by SEC-HPLC analysis. (a) Feedstock; (b) flow through fraction; (c) elution fraction; (d) CIP fraction. The mAb peak is at 16.6 min
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