title: Methods of preparation of recombinant cytokine proteins.
abstract: 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.
authors.affiliation: Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklikho-Maklaya 16/10, Moscow, GSP-7, 117871, Russia.
authors.forename: V E
authors.initials: VE
authors.lastname: Klyushnichenko
authors.forename: R V
authors.initials: RV
authors.lastname: Tikhonov
authors.forename: S E
authors.initials: SE
authors.lastname: Pechenov
authors.forename: S A
authors.initials: SA
authors.lastname: Yakimov
authors.forename: L N
authors.initials: LN
authors.lastname: Shingarova
authors.forename: V G
authors.initials: VG
authors.lastname: Korobko
authors.forename: A N
authors.initials: AN
authors.lastname: Wulfson
mesh_terms: 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*
chemical_list: D016207:Cytokines; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha