Evaluation of human interferon adsorption performance of Cibacron Blue F3GA attached cryogels and interferon purification by using FPLC system
Küçük Resim Yok
Tarih
2012
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
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 38.2 mg/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 mu g to 18 mu g, and interferon activity, from 2.6 x 10(3) IU to 2.2 x 10(3) IU, were determined, specific antiviral activity increased from 7.19 IU/mu g to 122.2 IU/mu g. 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. (C) 2012 Elsevier B.V. All rights reserved.
Açıklama
Anahtar Kelimeler
Poly(HEMA) cryogel, Cibacron Blue F3GA, Interferon, Affinity purification, Fast protein liquid chromatography
Kaynak
Journal of Chromatography B-Analytical Technologies in The Biomedical and Life Sciences
WoS Q Değeri
Q2
Scopus Q Değeri
Q2
Cilt
893
