期刊
ANALYTICA CHIMICA ACTA
卷 1017, 期 -, 页码 57-65出版社
ELSEVIER
DOI: 10.1016/j.aca.2018.02.012
关键词
Affinity monolith; Specific peptide ligand; Monoclonal antibody; Purification; Cell culture medium
资金
- National Natural Science Foundation of China [81673391, 81703460]
- China Post-doctoral Science Foundation [2016M602610, 2017T100662]
- Science and Technology Planning Project of Guangdong Province, China [2016A040403056]
The rapid development of monoclonal antibodies (mAbs) in therapeutic and diagnostic applications has necessitated the advancement of mAbs purification technologies. In this study, a biomimetic small peptide ligand 3,5-di-tert-butyl-4-hydroxybenzoic acid-Arg-Arg-Gly (DAAG) functionalized monolith was fabricated through a metal ion chelation-based multi-step approach. The resulting monolith showed good chromatographic performance. Compared with the Ni2+ based IMAC monolith, the DAAG functionalized monolith exhibited not only excellent specificity but also higher dynamic binding capacity (DBC). The 10% DBC and 50% DBC for hIgG reached as high values as 26.0 and 34.6 mg/mL, respectively, at a ligand density of 8.8 mmol/mL, due to the high porosity and accessibility of the monolithic matrix. Moreover, the stability of the DAAG functionalized monolith in successive breakthrough experiments indicates that it has a promising potential for long-term use in mAbs purification. Finally, the DAAG functionalized monolith was successfully applied to the purification of trastuzumab or human immunoglobulin G (hIgG) from biological samples. (C) 2018 Elsevier B.V. All rights reserved.
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