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Albumin separation with Cibacron Blue carrying macroporous chitosan and chitin affinity membranes
Institution:1. Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, MO 64108, USA;2. Department of Genetics, School of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA;3. ISTRA, Department of Chemistry, Abeda Inamdar Senior College, University of Pune, India;1. Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO 80045, USA;2. Structural Biology and Biochemistry Program, University of Colorado School of Medicine, Aurora, CO 80045, USA
Abstract:Cibacron Blue F3GA, Procion Red HE-3B and Procion Blue MX-R were immobilized on macroporous chitosan and chitin membranes with concentrations as high as 10–200 μmol/ml membrane. These dyed membranes were chemically and mechanically stable, could be reproducibly prepared, and operated at high flow rates. Human serum albumin (HSA) and bovine serum albumin (BSA) were selected as model proteins, and their adsorption on and desorption from the dyed chitosan membranes investigated. The Cibacron Blue F3GA membranes had a higher protein adsorption capacity, much greater for HSA than BSA, than the other dyed membranes. About 8.4 mg HSA/ml membrane were adsorbed at saturation by Cibacron Blue F3GA–chitosan membranes from a 0.05 M Tris–HCl/0.05 M NaCl, pH 8 solution. The chitin membranes had a lower dye content and hence a lower protein adsorption capacity than the chitosan membranes. The effects of important operation parameters (flow rate, protein concentration and loading) were also investigated. Cibacron Blue F3GA–chitosan membranes were employed for the separation of HSA from human plasma and high purity HSA thus obtained. This suggests that these membranes could be used for large-scale plasma fractionation.
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