title: Evaluation of human interferon adsorption performance of Cibacron Blue F3GA attached cryogels and interferon purification by using FPLC system.
abstract: In this study, we have focused our attention on preparing supermacroporous cryogels as a potential dye-affinity adsorbent for interferon purification. For this purpose, 2-hydroxyethyl methacrylate (HEMA) and Cibacron Blue F3GA (CB) were selected as main monomer and dye-ligand. Cibacron Blue F3GA attached supermacroporous poly(2-hydroxyethyl methacrylate) [poly(HEMA)/CB] cryogels were prepared and characterized by swelling test, scanning electron microscopy, elemental analysis, and FTIR. After that, the effecting factors such as pH, concentration, interaction time, and ionic strength on the interferon separation were evaluated. The maximum adsorption capacity of poly(HEMA)/CB cryogels was obtained as 38.2mg/g at pH 6.0. Fast protein liquid chromatography (FPLC) system was used for interferon purification from human gingival fibroblast extract. The chromatography parameters, capacity and selectivity factors, resolution and theoretical plate number were found as 7.79, 9.62, 4.23 and 554, respectively. Although some decreases in total protein content, from 320 μg to 18 μg, and interferon activity, from 2.6 × 10(3)IU to 2.2 × 10(3)IU, were determined, specific antiviral activity increased from 7.19 IU/μg to 122.2 IU/μg. The purified interferon samples have 97.6% purity determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After repeated ten adsorption-desorption cycles, no significant decrease was determined in adsorption capacity of cryogel. In result, poly(HEMA)/CB cryogels have an application potential for rapid, cheap and specific purification of interferon.
authors.affiliation: Inonu University, Faculty of Pharmacy, Biochemistry Division, Malatya, Turkey.
authors.forename: Ali
authors.initials: A
authors.lastname: Doğan
authors.forename: Serpil
authors.initials: S
authors.lastname: Özkara
authors.forename: Müfrettin Murat
authors.initials: MM
authors.lastname: Sarı
authors.forename: Lokman
authors.initials: L
authors.lastname: Uzun
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D059985:Cryogels / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016898:Interferon-alpha / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008689:Methacrylates / Q000737:chemistry; D009994:Osmolar Concentration; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014227:Triazines / Q000737:chemistry* / Q000378:metabolism
chemical_list: D059985:Cryogels; D016898:Interferon-alpha; D008689:Methacrylates; D011994:Recombinant Proteins; D014227:Triazines; C012842:Cibacron Blue F 3GA; C005044:hydroxyethyl methacrylate