An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme

Lu Wang, Rongsheng Zhang, Robert Eisenthal, John Hubble

Research output: Contribution to journalArticle

3 Citations (Scopus)

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)]
Original languageEnglish
Pages (from-to)37-42
Number of pages6
JournalBiotechnology and Applied Biochemistry
Volume45
Issue number1
Publication statusPublished - 2006

Fingerprint

Hydrogel
Muramidase
Hydrogels
Egg White
Enzymes
Recovery
Adsorption
Shielding
Dextran
Dextrans
Ligands
Gels
Ovalbumin
Adsorbates
Isotherms
Desorption
Buffers
Proteins

Keywords

  • shielded 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

Cite this

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

An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme. / Wang, Lu; Zhang, Rongsheng; Eisenthal, Robert; Hubble, John.

In: Biotechnology and Applied Biochemistry, Vol. 45, No. 1, 2006, p. 37-42.

Research output: Contribution to journalArticle

Wang, L, Zhang, R, Eisenthal, R & Hubble, J 2006, 'An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme', Biotechnology and Applied Biochemistry, vol. 45, no. 1, pp. 37-42.
Wang, Lu ; Zhang, Rongsheng ; Eisenthal, Robert ; Hubble, John. / An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme. In: Biotechnology and Applied Biochemistry. 2006 ; Vol. 45, No. 1. pp. 37-42.
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AB - 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)]

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