Preparation and characterization of mixed-mode magnetic adsorbent with p-amino-benzamidine ligand: Operated in a magnetically stabilized fluidized bed reactor for purification of trypsin from bovine pancreas

•The magnetic p(GMA-co-MMA) beads was synthesized and an affinity multi-modal ligand (i.e., p-amino-benzamidine) was immobilized.•The mp(GMA-co-MMA)-p-AMA affinity beads was characterized using FTIR, SEM and VSM.•Adsorption characteristics of trypsin on mp(GMA-co-MMA)-p-AMA affinity beads were well studied.•The affinity beads were used in a magnetically stabilized fluidized bed reactor to purify trypsin from crude pancreas extract.•The purification factor of trypsin from crude pancreas extract was 7.3 folds. The magnetic beads were synthesized using glycidylmethacrylate (GMA) and methylmethacrylate (MMA) monomers. A multimodal ligand (i.e., p-amino-benzamidine) was covalently immobilized onto magnetic beads after glutaraldehyde activation, and consequently used for purification of the trypsin from bovine pancreas. The p-amino-benzamidine ligand immobilized magnetic beads were characterized by FTIR, VSM, SEM, and analytical methods. Trypsin adsorption experiments were investigated under different experimental conditions (i.e., medium pH, initial trypsin concentration, temperature, and ionic strength) in a batch system. Maximum trypsin adsorption capacity was found to be 75.9±2.6mg/g beads. Adsorbed trypsin was eluted by using (0.1M acetate buffer, pH 3.0) with a 97% recovery. The purification factor of trypsin from crude pancreas extract was 8.7 folds. The purity of the eluted trypsin from p-amino-benzamidine functionalized magnetic beads was determined as 86% by HPLC. The method developed in this report was successfully applied for purification of the trypsin from crude pancreas extract in a magnetically stabilized fluidized bed reactor.