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An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme


Reference:

Wang, L., Zhang, R., Eisenthal, R. and Hubble, J., 2006. An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme. Biotechnology and Applied Biochemistry, 45 (1), pp. 37-42.

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Abstract

The present paper addresses the selective recovery of lysozyme from egg white using CM-dextran (carboxymethyldextran)-based hydrogels contg. Cibacron Blue as an affinity ligand and co-immobilized BSA intended to act as a shielding agent to reduce non-specific adsorption. Initial studies using pure lysozyme were conducted that indicated that the adsorption capacity increased with ligand d. and that adsorption was well described by a Langmuir-type isotherm. The inclusion of BSA as a putative shielding agent did not decrease the adsorption capacity for lysozyme in single-adsorbate expts. To assess the effectiveness of the shielding strategy, subsequent expts. were conducted with both defined lysozyme/ovalbumin mixts. and hen's-egg white. From these studies, the optimal operating conditions for lysozyme recovery have been detd. These include: optimal initial egg-white concn. [a 10% (vol./vol.) soln. of native egg white in the chosen buffer], affinity-ligand d. (1.86 mM) and ligand-to-shielding-agent ratio (4:1). The purity of lysozyme obtained from egg white was improved from 69% with a non-shielded hydrogel to 94% with an intrinsically shielded hydrogel. Finally, the possibility of using a protein, rather than dextran-backbone-based, hydrogel was investigated. It was found that BSA could take the place of CM-dextran as the gel backbone in a simplified synthesis, producing a gel which also proved effective for lysozyme recovery with a 30% lysozyme in egg-white soln. purified to approx. 92% in a single adsorption-desorption cycle. [on SciFinder (R)]

Details

Item Type Articles
CreatorsWang, L., Zhang, R., Eisenthal, R. and Hubble, J.
Uncontrolled Keywordsshielded hydrogel lysozyme purifn adsorption desorption, co-immobilized bovine, dissociation constant (intrinsically shielded hydrogel for the adsorptive recovery of lysozyme), uses (uses), albumins role, ract (reactant or reagent) (serum, unclassified), intrinsically shielded hydrogel for the adsorptive recovery of lysozyme), hydrogels, purification (affinity, proc (process) (displacement by lysozyme, egg white (of hen egg white, proc (process), rct (reactant), ovalbumin role, pep (physical, adsorption (protein, shielding agent, immobilization (enzyme, engineering or chemical process), nuu (other use
DepartmentsFaculty of Engineering & Design > Chemical Engineering
RefereedYes
StatusPublished
ID Code853

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