NDLI: Thiacarbocyanine as ligand in dye-affinity chromatography for protein purification. II. Dynamic binding capacity using lysozyme as a model.

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Thiacarbocyanine as ligand in dye-affinity chromatography for protein purification. II. Dynamic binding capacity using lysozyme as a model.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Boto, R. E. F. Anyanwu, U. Sousa, F. Almeida, P. Queiroz, J. A.
Description	Country affiliation: Portugal Author Affiliation: Boto RE ( Departamento de Química e Unidade de I & D de Materiais Têxteis e Papeleiros, Universidade da Beira Interior, Covilhã, Portugal.)
Abstract	A constant development of dye-affinity chromatography to replace more traditional techniques is verified, with the aim of increasing specificity in the purification of biomolecules. The establishment of a new dye-affinity chromatographic support imposes their complete characterization, namely with relation to the binding capacity for proteins, in order to evaluate its applicability on global purification processes. Following previous studies, the adsorption of lysozyme onto a thiacarbocyanine dye immobilized on beaded cellulose was investigated. The effect of different parameters, such as temperature, ionic strength, pH, protein concentration and flow rate, on the dynamic binding capacity of the support to retain lysozyme was also studied. Increasing the temperature and the lysozyme concentration had a positive effect on the dynamic binding capacity (DBC), whereas increasing the ionic strength and the flow rate resulted in the opposite. It was also discovered that the pH used had an important impact on the lysozyme binding onto the immobilized dye. The maximum DBC value obtained for lysozyme was 8.6 mg/mL, which was achieved at 30 degrees C and pH 9 with a protein concentration of 0.5 mg/mL and a flow rate of 0.05 mL/min. The dissociation constant (K(d)) obtained was 2.61 +/- 0.36 x 10(-5 )m, proving the affinity interaction between the thiacarbocyanine dye ligand and the lysozyme.
File Format	HTM / HTML
ISSN	02693879
Issue Number	9
Volume Number	23
e-ISSN	10990801
Journal	Biomedical Chromatography
Language	English
Publisher	Wiley
Publisher Date	2009-09-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Analytical Chemistry Discipline Molecular Biology Discipline Biochemistry Discipline Pharmacology Carbocyanines Chemistry Chromatography, Affinity Methods Coloring Agents Muramidase Adsorption Metabolism Hydrogen-ion Concentration Linear Models Models, Chemical Isolation & Purification Osmolar Concentration Protein Binding Temperature Journal Article
Content Type	Text
Resource Type	Article
Subject	Medicine Analytical Chemistry Drug Discovery Clinical Biochemistry Biochemistry Molecular Biology Pharmacology

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