title: Composite cryogels for lysozyme purification.
abstract: Beads-embedded novel composite cryogel was synthesized to purify lysozyme (Lyz) from chicken egg white. The poly(hydroxyethyl methacrylate-N-methacryloyl-L-phenylalanine) (PHEMAPA) beads of smaller than 5 µm size were synthesized by suspension polymerization and then embedded into a poly(hydroxyethyl methacrylate) (PHEMA)-based cryogel column. The PHEMAPA bead-embedded cryogel (BEC) column was characterized by swelling tests, scanning electron microscopy (SEM), surface area measurements by the Brunauer-Emmett-Teller (BET) method, elemental analysis, and flow dynamics. The specific surface area of the PHEMAPA BEC was found as 41.2 m(2) /g using BET measurements. Lyz-binding experiments were performed using aqueous solutions in different conditions such as initial Lyz concentration, pH, flow rate, temperature, and NaCl concentration of an aqueous medium. The PHEMAPA BEC column could be used after 10 adsorption-desorption studies without any significant loss in adsorption capacity of Lyz. The PHEMAPA BEC column was used to purify Lyz from chicken egg white, and gel electrophoresis was used to estimate the purity of Lyz. The chromatographic application of the PHEMAPA BEC column was also performed using fast protein liquid chromatography.
authors.affiliation: Biochemistry Division, Department of Chemistry, Hacettepe University, Ankara, Turkey.
authors.forename: Gözde
authors.initials: G
authors.lastname: Baydemir
authors.forename: Emir Alper
authors.initials: EA
authors.lastname: Türkoğlu
authors.forename: Müge
authors.initials: M
authors.lastname: Andaç
authors.forename: Işık
authors.initials: I
authors.lastname: Perçin
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000818:Animals; D002642:Chick Embryo; D059985:Cryogels / Q000737:chemistry*; D004527:Egg Proteins; D004529:Egg White; D004789:Enzyme Activation; D004795:Enzyme Stability; D059625:Liquid-Liquid Extraction / Q000379:methods*; D009113:Muramidase / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D059985:Cryogels; D004527:Egg Proteins; D009113:Muramidase
keywords: bead-embedded cryogels; hydrophobic affinity chromatography (HIC); lysozyme