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 |
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| Institution: | 1. Biochemical Processing and Biomaterial Research Laboratory, Gazi University, Teknikokullar, 06500 Ankara, Turkey;2. Department of Chemistry, Faculty of Sciences, Gazi University, Teknikokullar, 06500 Ankara, Turkey;3. School of Medicine, Istanbul Kemerburgaz University, 34217 Istanbul, Turkey;1. Centro Universitário Estácio do Ceará Via Corpus Rua Eliseu Uchoa Becco, n° 600 – Bairro Água Fria, CEP: 60810-270, Fortaleza, Ceará, Brazil;2. Departmento de Bioquímica e Biologia Molecular/UFC, Campus do Pici, Caixa Postal 6033, 60451-970 Ceará, Brazil;3. Departmento de Química Orgânica e Inorgânica/UFC, Campus do Pici, Caixa Postal 6021, 60455-760 Ceará, Brazil;4. Departmento de Fisiologia e Farmacologia/UFC, Coronel Nunes de Melo, 1127 Rodolfo Teófilo, 60430-270 Ceará, Brazil;1. School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, Liaoning, People''s Republic of China;2. Jiangshu Feixiang Chemical Company Limited Duty, Suzhou 215613, Jiangshu, People''s Republic of China;1. Laboratory for Biophysics of Macromolecular Aggregates, Department of Biomedical Engineering, Wroc?aw University of Science and Technology, Wybrze?e Stanis?awa Wyspiańskiego 27, 50-370 Wroc?aw, Poland;2. Research and Development Centre, Specialized Hospital in Wroc?aw, Kamieńskiego 73 A, 51-124 Wroc?aw, Poland;1. Argentine Institute of Mathematics (I. A. M.), National Research Council (CONICET), Buenos Aires 1083, Argentina;2. Collegium Basilea – Institute for Advanced Study, Basel CH4053, Switzerland |
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| Abstract: | 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 ± 2.6 mg/g beads. Adsorbed trypsin was eluted by using (0.1 M 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. |
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| Keywords: | Magnetic beads Affinity chromatography Trypsin Fluidized bed reactor |
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