Biomimetic small peptide functionalized affinity monoliths for monoclonal antibody purification

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 μmol/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. [Display omitted] •Biomimetic small peptide functionalized monolith was fabricated through a metal ion chelation-based multi-step approach.•The poly(GMA-co-EDMA) monolith was used as the support material.•The DAAG functionalized monolithic column exhibits not only higher specificity and dynamic binding capacity, but also lower non-specificity than the Ni2+ based IMAC monolith for the purification of mAbs.•The column shows a great application potential for mAbs capture and purification from cell culture medium.