Methods of preparation of recombinant cytokine proteins.
Two schemes for efficient and productive isolation for mutant human recombinant tumor necrosis factor-alpha (TNF-alpha R32H) from Escherichia coli cells were developed. The methods include membrane filtration, ion-exchange chromatography and gel filtration, and centrifugation with subsequent free-flow electrophoresis as an alternative procedure. The target product was obtained as active trimer with total yield more than 50% and greater than 98% purity according to PAGE, size-exclusion chromatography, HPLC, and HPCE.
Klyushnichenko
V E
VE
Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklikho-Maklaya 16/10, Moscow, GSP-7, 117871, Russia.
Tikhonov
R V
RV
Pechenov
S E
SE
Yakimov
S A
SA
Shingarova
L N
LN
Korobko
V G
VG
Wulfson
A N
AN
D002845:Chromatography / Q000379:methods; D016207:Cytokines / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D009154:Mutation / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification*; D017421:Sequence Analysis; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000302:isolation & purification*
D016207:Cytokines; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha