Adsorption of papain with Cibacron Blue F3GA carrying chitosan-coated nylon affinity membranes.
Covalent coupling of chitosan (CS) to activated nylon membrane was performed after the reaction of the microporous nylon membrane with formaldehyde. Non-specific adsorption on the CS-coated nylon membrane decreased greatly, compared with plain nylon membrane. The dye Cibacron Blue F3GA (CB F3GA) as a ligand was then covalently immobilized on the CS-coated membranes. Physical properties of the composite membrane and its applications in affinity membrane chromatography were examined. The contents of CS and CB F3GA-attached membranes were 89.6 mg/g nylon membrane and 146.1 micromol/g nylon membrane, respectively. These CB F3GA-attached composite membranes were used in the papain adsorption studies. Higher papain adsorption capacity, up to 235.3mg/g affinity membrane, was obtained. The adsorption isotherm fitted the Freundlich model well. Significant amount of the adsorbed papain (about 94.3%) was eluted by 1.0M NaSCN at pH 9.0. Experiments on regeneration and dynamic adsorption were also performed. It appears that CB F3GA-CS nylon membranes can be applied for papain separation without causing any denaturation.
Nie
Hua-Li
HL
Institute of Biological Sciences & Biotechnology, Donghua University, Shanghai 201620, China.
Zhu
Li-Min
LM
D000327:Adsorption; D048271:Chitosan / Q000737:chemistry*; D008567:Membranes, Artificial; D009757:Nylons / Q000737:chemistry*; D010206:Papain / Q000737:chemistry / Q000302:isolation & purification*; D014227:Triazines / Q000737:chemistry*
D008567:Membranes, Artificial; D009757:Nylons; D014227:Triazines; C012842:Cibacron Blue F 3GA; D048271:Chitosan; D010206:Papain