title: Purification of recombinant human tumor necrosis factor precursor from Escherichia coli.
abstract: To study its biological functions, tumor necrosis factor precursor (proTNF) with a molecular size of 26-KDa was obtained as a recombinant protein from Escherichia coli. The recombinant proTNF was successfully accumulated in the insoluble form, corresponding to about 10-15% of total E. coli proteins. Solubilization, gel filtration and anion exchange chromatography were performed under denatured conditions followed by dialysis in phosphate-buffered saline. These processes removed most of the contaminating bacterial proteins, yielding proTNF with a purity of about 70-80%. This recombinant proTNF is expected to be useful for functional studies on activated macrophages with membrane integrated proTNF.
authors.affiliation: Biotechnology Research Center, Teikyo University, Kawasaki 2, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Tanabe
authors.forename: K
authors.initials: K
authors.lastname: Noguchi
authors.forename: A
authors.initials: A
authors.lastname: Morikawa
authors.forename: D
authors.initials: D
authors.lastname: Mizuno
authors.forename: G
authors.initials: G
authors.lastname: Soma
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D008970:Molecular Weight; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011498:Protein Precursors; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C061153:tumor necrosis factor precursor