title: Dye-ligand affinity chromatography on continuous beds.
abstract: Continuous beds derivatized with three triazine dyes (Cibacron Blue 3G-A, Pricon Red HE-3B and Pricon Red H-3B) were used for chromatographic purification of dehydrogenases from yeast enzyme concentrate. All three columns, which were prepared by a very simple and cost-effective method, provided strong binding of glucose-6-phosphate dehydrogenase and lactate dehydrogenase. The Cibacron Blue 3G-A column showed high affinity for alcohol dehydrogenase. Under the same conditions, the Pricon Red HE-3B column showed a lower affinity and the Pricon Red H-3B column showed none. The adsorbed dehydrogenases were eluted specifically from the columns in high yields (71-113% by desorption with the coenzymes NADP, NADH and NAD respectively). Non-specific binding of human serum albumin and transferrin to these columns was also investigated. Enzyme assays and analyses by capillary electrophoresis showed that the continuous beds derivatized with triazine dyes gave a high degree of purification.
authors.affiliation: Department of Biochemistry, University of Uppsala, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Mohammad
authors.forename: A
authors.initials: A
authors.lastname: Zeerak
authors.forename: S
authors.initials: S
authors.lastname: Hjertén
mesh_terms: D000426:Alcohol Dehydrogenase / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D004396:Coloring Agents; D004586:Electrophoresis / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification; D008024:Ligands; D013056:Spectrophotometry, Ultraviolet
chemical_list: D004396:Coloring Agents; D008024:Ligands; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase