\Integration of Expanded Bed Adsorption and Hydrophobic Charge-Induction Chromatography for Monoclonal Antibody Separation

The integration of expanded bed adsorption (EBA) with hydrophobic charge-induction chromatography (HCIC) affords a promising new technology to capture antibody from the complex feedstock. New EBA resin T-ABI was prepared with hydrophobic charge-induction ligand 5-aminobenzimidazole (ABI) and used to separate monoclonal antibody (mAb) from CHO cell culture broth. The static and dynamic adsorption behaviors of hIgG and mAb were investigated, and the typical properties of pH dependence and salt tolerance were found. High dynamic binding capacities at 10% breakthrough (18.1-21.4 mg/mL resin) were obtained even for high operation velocities (711-1203 cm/h) in an expanded bed. By optimization of loading pH, elution pH, and expansion factor, hIgG could be separated from the protein mixture (2 mg/mL hIgG and. 10 mg/mL bovine serum albumin) with high efficiency. Finally, mAb was separated directly from the CHO cell culture broth with T-ABI EBA under optimized conditions (loading at pH 7.0, elution at pH 4.0 or 4.5, expansion factor of 2.0), and the purity reached 93.7-97.7% with the recovery of 72.6-79.4%. The results indicated that T-ABI resin was suitable to capture hIgG from the complicated feedstock. New separation process with T-ABI EBA showed a promising potential for antibody purification with high productivity and process efficiency, which combined the advantages of HCIC and EBA.