title: Affinity adsorption of lysozyme on a macroligand prepared with Cibacron Blue 3GA attached to yeast cells.
abstract: The objective of this study was the development of affinity adsorbent particles with the appropriate characteristics to be applied in protein purification using the affinity ultrafiltration method. To prepare affinity macroligands Cibacron Blue 3GA, as a ligand molecule, was immobilized by covalent bonding onto yeast cell walls, the support material or matrix. The maximum attachment of the ligand to the matrix was 212 μmol/g (ligand dry weight/yeast dry weight). Lysozyme was selected as the protein model for the adsorption studies. Its adsorption onto the matrix without ligand and matrix with attached ligand were investigated batch-wise. The adsorption equilibrium isotherms appeared to follow a typical Langmuir isotherm. The maximum adsorption capacity (q(m)) of the Cell-Cibacron macroligand for lysozyme was 110 mg/ml of wet macroligand. The adsorbent was also employed for the separation of lysozyme from hen egg white. High purity lysozyme was obtained.
authors.affiliation: Laboratory of Molecular Biology and Biotechnology, Department of Biochemistry, San Luis University, Ejército de los Andes 950, D5700HHW, San Luis, Argentina. ferraris@unsl.edu.ar
authors.forename: María
authors.initials: M
authors.lastname: del Pilar Ferraris
authors.forename: Guillermo I
authors.initials: GI
authors.lastname: Barrera
authors.forename: A Pérez
authors.initials: AP
authors.lastname: Padilla
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Rodríguez
mesh_terms: D000327:Adsorption; D018914:Cells, Immobilized / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D014227:Triazines / Q000737:chemistry*
chemical_list: D014227:Triazines; C012842:Cibacron Blue F 3GA; D009113:Muramidase