title: [Isolation of expressed in E. coli human interferon beta1b (Ser17) by ion-exchange chromatography].
abstract: A method for isolation of interferon beta1b (Serl7) from inclusion bodies, comprising the steps of solution and reduction of protein from the inclusion bodies, refolding, chromatography on DEAE-Sepharose, chromatography on SP-Sepharose, concentrating, desalting and addition of stabilizers. The solution of reduced protein was diluted with pH 8.0 buffer of 50 mM Tris-HCl, 25 microM CuCl2 and 0.5% Twin 20 for refolding. We used gradient of pH (from 9.3 upto 11.3) for elution of interferon-beta from cation-exchange column. We concentrated of eluate and then desalted on the Sephadex G-50 column with 1 mM NaOH. Then the protein solution was neutralized with mannitol and Na-phosphate. Obtained preparation of interferon-beta was pure by gel-electrophoresis and by HPLC analysis, and had practically indentical level of antiproliferative activity with well-known preparation of Betaferone. Thus we show the possibility of isolation and obtaining of pure and active interferone-beta by ion-exchange chromatography in the presence of non-ion detergent Twin 20. We believe this method for interferon betalb preparation is perspective for scaling and using in the develop of industrial technology for production of this preparation.
authors: V P
authors: VP
authors: Romanov
authors: V V
authors: VV
authors: Bezuglov
authors: M Iu
authors: MIu
authors: Bobrov
authors: T I
authors: TI
authors: Kostromina
authors: S A
authors: SA
authors: Feofanov
authors: A I
authors: AI
authors: Miroshnikov
mesh_terms: D002852:Chromatography, Ion Exchange / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D000068576:Interferon beta-1b; D016899:Interferon-beta / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D000068576:Interferon beta-1b; D016899:Interferon-beta