Rapid purification method for human recombinant tumor necrosis factor alpha.
Human recombinant tumor necrosis factor alpha was purified in a single step to about 95% purity from Escherichia coli lysate by chromatography on hydroxyapatite. The last traces of contaminants were removed by fast protein liquid chromatography on a Mono Q column. The final product was found to be pure by gel electrophoresis with silver staining. A molecular mass of approximately 17,000 and a specific activity of 4.3 x 10(6) U/mg after a single purification step were found.
Department of Biochemistry, Faculty of Medicine, Université Laval, Sainte-Foy, Québec, Canada.
A
A
Paquet
A
A
Lévesque
M
M
Pagé
D000596:Amino Acids / Q000032:analysis; D000837:Anion Exchange Resins; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D006882:Hydroxyapatites; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification; D012117:Resins, Synthetic; D013056:Spectrophotometry, Ultraviolet; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000302:isolation & purification*
D000596:Amino Acids; D000837:Anion Exchange Resins; D006882:Hydroxyapatites; D011994:Recombinant Proteins; D012117:Resins, Synthetic; D014409:Tumor Necrosis Factor-alpha; C042855:Mono Q