Affinity adsortion of human interferon-?

Interferons are biological active proteins which are synthesized by leukocytes, lenfoblastoid and myeloblastoid cell lines. Especially, they have been used in treatments of kidney cancer, malignant melonoma, multiple myeloma, carsinoid, lymphoma and leukemia. In addition, they are also used in treatment AIDS, chronic hepatitis B and C. Cryogels are gel matrix which are prepared by using partially frozen monomer and polymer solutions. Generally, cryogels have interconnected macropores. Therefore, cryogels can be used in wide scale range from nano to micro without any diffusion problem.In present study, dye attached monolithic cryogels, potential adsorbent for bioaffinity chromatography, were prepared for interferon-? purification, then, optimal purification conditions and interferon purification performance of the adsorbent were investigated. At first, poly(2-hydroxyethyl methacrylate) cryogels were prepared. Then, Cibacron Blue F3GA, dye ligand, was attached onto cryogels. Cryogels were characterized by swelling test, scanning electron microscopy, elemental analysis, surface area measurements and FTIR. After that, interferon adsorption studies were carried out from aqueous solutions and natural interferon source, human gingival fibroblast. In order to determine the optimal adsorption conditions, pH, concentration, interaction time, and ionic strength were investigated. The maximum adsorption capacity of Cibacron Blue F3GA attached poly(HEMA) cryogels was obtained as 38.2 mg/g at pH 6.0 and 2 h incubation. Interferon purification from natural source was carried out by using fast protein liquid chromatography system. The chromatography parameters, capacity and selectiviy factors, resolution and theoritical plate number were found as 7.79, 9.62, 4.23 and 554, respectively. Interferon desorption from Cibacron Blue F3GA attached poly(HEMA) cryogels was done by using 1M NaCl solution. After ten times repeated adsorption-desorption cycles, no significant decrease could not be determined in adsorption capacity of cryogel. In result, Cibacron Blue F3GA attached poly(HEMA) cryogels have an application potential for rapid, cheap and specific purification of interferon.