title: Purification of human tumour necrosis factor by membrane chromatography.
abstract: The recombinant human tumour necrosis factor alpha from an extract of Escherichia coli was enriched to homogeneity according to specific activity and sodium dodecyl sulphate-polyacrylamide gel electrophoresis by purification using anion-exchange HPLC and hydrophobic interaction HPLC. Parallel experiments with the same separation methods, but carried out with membrane chromatography on compact discs, gave similar results in terms of yield and purity of the product. The active form of the protein is a trimer. The second isolation step, hydrophobic interaction chromatography, causes dissociation of the trimer into monomers and a partial loss of the biological activity of the protein. The phenomenon occurs on both the column and the disc. This in turn indicates strongly that the dissociation of the protein is a consequence of interaction between the samples and the hydrophobic ligand, and is not caused by non-specific interaction with the matrix.
authors.affiliation: Lek, Pharmaceutical and Chemical Company, Research and Development, Ljubljana Slovenia.
authors.forename: J
authors.initials: J
authors.lastname: Luksa
authors.forename: V
authors.initials: V
authors.lastname: Menart
authors.forename: S
authors.initials: S
authors.lastname: Milicic
authors.forename: B
authors.initials: B
authors.lastname: Kus
authors.forename: V
authors.initials: V
authors.lastname: Gaberc-Porekar
authors.forename: D
authors.initials: D
authors.lastname: Josic
mesh_terms: D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha