The study of adsorption characteristics of lysozyme onto dye affinity nanofibrous membrane

• Main Authors: Chi-Lin Yang, 楊奇霖
• Other Authors: Yu-Kaung Chang
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/t254z7

碩士 === 明志科技大學 === 化學工程系生化工程碩士班 === 103 === Polyacrylonitrile (PAN) nanofiber membrane was prepared by electrospinning technique. After heat treatment and alkaline hydrolysis, the ion exchange membrane (namely P-COOH) was grafted with ethylenediamine (EDA) and chitosan molecule, respectively. The obtained P-NH2 and P-Chitosan membranes were then covalently immobilized with Reactive Blue 49 dye, respectively to be used as dye ligand affinity membranes. The two dyed membranes were characterized in terms of fiber diameter, porosity, pore size, ligand density and binding capacities. The membrane was applied to evaluate the binding capacity of a model protein, lysozyme, under various operating parameters (e.g., adsorption pH, EDA, chitosan and dye concentrations, ionic strength, and temperature) were studied in batch modes. Under these circumstances, the optimal synthesis of dyed membrane and adsorption conditions for lysozyme could be obtained. The dynamic adsorption characteristics of these two dyed membranes for lysozyme using membrane chromatography were assessed by measurements of the breakthrough curves. The influences of operating conditions (e.g., adsorption pH, lysozyme concentration, and flow rate) on the adsorption performance of membrane were investigated in an ÄKTA prime prime chromatographic system (GE Healthcare). On the basis of the experiments, the adsorption capacity of P-NH2-dye membrane for lysozyme was higher as compared to the P-Chitosan-dye membrane. Moreover, the one-step elution scheme (1.0 M NaCl, pH 4) was chosen for eluting the adsorbed lysozyme. The lysozyme fraction eluted by 1.0 M NaCl for P-NH2-dye and P-Chitosan-dye membrane was recovered with a yield of 99% and 97%, respectively at a flow rate of 0.1 ml/min.