Rapid purification of recombinant human tumor necrosis factor beta.
A rapid and improved method for the purification of recombinant human tumor necrosis factor beta (rhTNF-beta) from Escherichia coli HB 101 cells has been developed. The method utilized sequential steps of polyethylenimine (PEI) and ammonium sulfate precipitation to remove most of the extraneous proteins and nucleic acids from the cell extracts. The final step of purification consisted of DEAE-Sepharose chromatography at pH 7.5 in which rhTNF-beta was eluted with starting buffer. This procedure, when compared to the earlier methods of purification, is highly efficient since we could increase the overall yield of rhTNF-beta and reduce the purification time considerably. The final yield that we obtained from 1 liter of fermentation broth (containing approximately 80 g of wet cells) was 40-50 mg.
Loh
K C
KC
Bioprocessing Technology Unit, National University of Singapore.
Yao
Z J
ZJ
Yap
M G
MG
Chung
M C
MC
D000595:Amino Acid Sequence; D000645:Ammonium Sulfate; D000818:Animals; D011232:Chemical Precipitation; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006801:Humans; D007739:L Cells / Q000187:drug effects; D008233:Lymphotoxin-alpha / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D011094:Polyethyleneimine; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology
D008233:Lymphotoxin-alpha; D011993:Recombinant Fusion Proteins; D011094:Polyethyleneimine; D000645:Ammonium Sulfate