Recently, immunoglobulin G (IgG) has attracted great attentions in clinical medicine, biotechnology, stimulated the development of downstream purification technology of monoclonal antibodies. Now, the protein based affinity ligands during traditional chromatographic purification of IgG exist drawbacks including the high cost and low stability, which leads to the secondary contamination and biological toxicity to purified sample. Also the chromatographic agarose microspheres present shortcomings such as the gel compressibility and the mass transfer limitations. In this work, biomimetic peptide as the protein based ligands alternative, grafting on the supermacroporous dextran-poly(glycidyl methacrylate) microspheres (Dextran-PGMA) to overcome above drawbacks were prepared and to efficiently improve the IgG chromatographic separation. The hydroxy on dextran-grafted polymer chromatographic microspheres were converted to the epoxy groups in epichlorohydrin solution with 2 mol/L NaOH, and the epoxy groups were opened and coupled with biomimetic peptide, FYEILCH, to achieve the purpose of affinity modification. The SEM observed that the pore structure of macroporous was mostly maintained after coupling with the biomimetic peptide. The change of 10% dynamic binding capacity (DBC10%) of FYEILHC based on Dextran-PGMA microspheres and agarose microspheres under different flow rate (92~923 cm/h) was compared. The DBC10% of dextran-PGMA was declined only 8% under 923 cm/h, whereas the DBC10% of agarose was sharply declined about 25%. This phenomenon might be related to better mass transfer for IgG at higher flow rate on the Dextran-PGMA microspheres. The DBC10% of Dextran-PGMA microspheres maintained (21±1) mg/mL after cleaning-in-place (CIP) 40 cycles using 0.1 mol/L NaOH, indicating that the affinity medium had good chemical stability. The purity of recovered antibodies in plasma was 95.0%, demonstrating that this biomimetic peptide based affinity medium had great potential to purify IgG from complex biological samples. This biomimetic peptide grafted supermacroporous poly(glycidyl methacrylate) microspheres could meet the requirement for rapid and high-throughput separation of monoclonal antibody.
Jia GE Xiangming NA Xuexing WU Weixing YANG Dongxia HAO Guanghui MA
. Preparation of biomimetic peptide attached supermacroporous poly(glycidyl methacrylate) microspheres for monoclonal antibody purification and its performance[J]. The Chinese Journal of Process Engineering, 2020
, 20(1)
: 84
-90
.
DOI: 10.12034/j.issn.1009-606X.219166
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