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Related Experiment Video

Updated: Jul 15, 2026

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
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Lysozyme purification with dye-affinity beads under magnetic field.

Nilgün Başar1, Lokman Uzun, Ali Güner

  • 1Department of Chemistry, Hacettepe University, Ankara, Turkey.

International Journal of Biological Macromolecules
|April 10, 2007
PubMed
Summary

Magnetic poly(2-hydroxyethyl methacrylate) beads functionalized with Cibacron Blue F3GA efficiently purify lysozyme. These reusable beads demonstrate high adsorption capacity and enable significant recovery of pure lysozyme from egg white.

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Area of Science:

  • Biomaterials Science
  • Affinity Chromatography
  • Protein Purification

Background:

  • Poly(2-hydroxyethyl methacrylate) (PHEMA) is a versatile polymer with tunable properties.
  • Magnetic nanoparticles (Fe3O4) offer advantages for separation and recovery.
  • Cibacron Blue F3GA is a well-established ligand for albumin and related protein purification.

Purpose of the Study:

  • To synthesize and characterize magnetic PHEMA beads (mPHEMA) functionalized with Cibacron Blue F3GA.
  • To evaluate the adsorption capacity and reusability of these beads for lysozyme purification.
  • To assess the efficiency of mPHEMA/Cibacron Blue F3GA beads in purifying lysozyme from a complex biological matrix (egg white).

Main Methods:

  • Suspension polymerization of HEMA in the presence of Fe3O4 nanoparticles to create mPHEMA beads.

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  • Functionalization of mPHEMA beads with Cibacron Blue F3GA.
  • Characterization using FTIR and Raman spectroscopy.
  • Lysozyme adsorption studies in a magnetically stabilized fluidized bed under various conditions (pH, temperature, ionic strength, etc.).
  • Lysozyme purification from egg white and purity/activity assessment.

    • Main Results:

    • Synthesized spherical mPHEMA beads (80-120 µm) with a specific surface area of 56.0 m²/g.
    • mPHEMA/Cibacron Blue F3GA beads exhibited a high lysozyme adsorption capacity of 342 mg/g, significantly higher than unmodified mPHEMA beads (0.8 mg/g).
    • The beads maintained their adsorption capacity over 20 adsorption-desorption cycles.
    • Purification of lysozyme from egg white yielded a purity of 87.4% with a recovery of 79.6% and high specific activity (41.586 U/mg).

    Conclusions:

    • Magnetic mPHEMA beads functionalized with Cibacron Blue F3GA are effective for high-capacity lysozyme adsorption and purification.
    • The magnetic properties facilitate easy separation and recovery, while the dye ligand ensures specific binding.
    • These reusable beads offer a promising and efficient method for purifying lysozyme from crude sources like egg white.