Purification of lysozyme by multistage affinity filtration

Area of Science:

• Biochemistry
• Chemical Engineering
• Separation Science

Background:

• Protein purification is crucial for research and biotechnology.
• Existing methods may lack selectivity and efficiency for specific protein recovery.
• Developing advanced separation techniques is essential for isolating target proteins.

Purpose of the Study:

• To develop and evaluate a multistage affinity filtration process for selective protein purification.
• To optimize the recovery yield of lysozyme using Cibacron Blue 3GA immobilized on TSK gel.
• To model and analyze the protein adsorption and elution dynamics in the affinity filtration system.

Main Methods:

• Immobilization of Cibacron Blue 3GA onto TSK gel HW-65F to create an affinity adsorbent.
• Adsorption equilibrium studies to determine binding affinities for lysozyme and bovine serum albumin (BSA).
• Implementation of a three-stage affinity filtration system for protein separation.
• Development of a mathematical model incorporating diffusion parameters for system analysis.

Main Results:

• The developed affinity adsorbent exhibited high affinity for lysozyme and weaker binding for BSA.
• The three-stage affinity filtration system successfully purified lysozyme from both model mixtures and chicken egg white.
• Recovery yield of lysozyme from chicken egg white increased from 61% (one-stage) to 96% (three-stage).
• Experimental and modeling data validated the effectiveness of the multistage approach.

Conclusions:

• Multistage affinity filtration is an effective technique for selective protein recovery.
• The developed process significantly enhances lysozyme yield and purity.
• This method offers a promising approach for protein purification in various applications.