Affinity adsorption of lysozyme on a macroligand prepared with Cibacron Blue 3GA attached to yeast cells

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2011-09, Vol.879 (26), p.2741-2745
• Main Authors: del Pilar Ferraris, María, Barrera, Guillermo I., Padilla, A. Pérez, Rodríguez, Jorge A.
• Format: Article
• Language: English
• Subjects: adsorbents; Adsorption; Affinity macroligand; Affinity ultrafiltration; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; cell walls; Cells, Immobilized - chemistry; Cells, Immobilized - cytology; Cells, Immobilized - metabolism; chemical bonding; chromatography; Chromatography, Affinity - methods; Cibacron Blue 3GA; egg albumen; Electrophoresis, Polyacrylamide Gel; Fundamental and applied biological sciences. Psychology; General pharmacology; hens; Lysozyme; Medical sciences; Muramidase - chemistry; Muramidase - isolation & purification; Muramidase - metabolism; Pharmacology. Drug treatments; Protein Binding; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - metabolism; sorption isotherms; Triazines - chemistry; ultrafiltration; Yeast cells; yeasts
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2011.07.040

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.