Improved hollow-fibre membranes for dye-affinity chromatography.
Hollow-fibre membranes with different degrees of surface hydrophilicity were obtained by grafting mixtures of glycidyl methacrylate (GMA) and dimethyl acrylamide (DMAA) in various proportions, and Cibacron Blue F3G-A was attached to them through ammonia or glucamine spacers. Membrane hydrophilicity increased with the amount of dimethyl acrylamide in the grafted polymer. As the hydrophilicity increased the permeability decreased from 352 mL/cm2 min MPa for membranes grafted with GMA with ammonia spacer to 12.7 mL/cm2 min MPa for membranes grafted with GMA/DMAA 1/3 with glucamine spacer. Membranes grafted with GMA/DMAA 1/3 with ammonia spacer showed the best performance for BSA and lysozyme adsorption: maximum capacity was 15.3 +/- 2.2 mg BSA/mL membrane and 58.3 +/- 6.6 mg lysozyme/mL membrane while dissociation constants were 0.27 +/- 0.16 and 0.13 +/- 0.12 mg/mL, respectively. Over 80% of adsorbed proteins could be eluted with 2 M NaCl + 20% isopropanol in 20 mM sodium phosphate buffer, pH 7.0.
Wolman
Federico J
FJ
Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica (UBA), Junín 956 (1113) Buenos Aires, Argentina.
Smolko
Eduardo E
EE
Cascone
Osvaldo
O
Grasselli
Mariano
M
D000178:Acrylamides; D000327:Adsorption; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents; D004852:Epoxy Compounds; D057927:Hydrophobic and Hydrophilic Interactions; D008567:Membranes, Artificial; D008689:Methacrylates; D009113:Muramidase / Q000302:isolation & purification; D010539:Permeability; D011108:Polymers; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D013499:Surface Properties; D014227:Triazines
D000178:Acrylamides; D004396:Coloring Agents; D004852:Epoxy Compounds; D008567:Membranes, Artificial; D008689:Methacrylates; D011108:Polymers; D011506:Proteins; D014227:Triazines; D012710:Serum Albumin, Bovine; C012842:Cibacron Blue F 3GA; C099046:N,N-dimethylacrylamide; D009113:Muramidase; C007870:glycidyl methacrylate