Lysozyme, a naturally occurring enzyme abundant in chicken egg white (CEW), has gained significant industrial interest due to its antimicrobial and preservative properties. Its ability to hydrolyze the 1,4-β-linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in bacterial cell walls makes it effective against Gram-positive bacteria. Despite its importance, traditional purification methods such as ultrafiltration, precipitation, and packed-bed column chromatography often suffer from low efficiency, high operational cost, and lengthy processing times. To overcome these limitations, membrane chromatography has emerged as a promising alternative. This study focuses on the development and optimization of a polyacid ion exchange (IEX) nanofiber membrane for high-efficiency lysozyme recovery from CEW.
The polyacid IEX nanofiber membrane (P-BrA) was fabricated via electrospinning of polyacrylonitrile (PAN) followed by surface functionalization using ethylene diamine (EDA) and bromoacetic acid (BrA).NCAM-L1 Antibody In Vitro The resulting membrane exhibited a highly porous, interconnected fibrous structure with an average fiber diameter of 467.3 nm. Fourier-transform infrared spectroscopy confirmed successful grafting of amine and carboxyl groups, indicating chemical modification. Thermogravimetric analysis revealed good thermal stability up to 320°C, making the membrane suitable for ambient temperature operations. The membrane’s porosity was measured at 83.67%, with a mean pore size of 457.1 nm, facilitating efficient mass transfer.
Batch adsorption experiments were conducted to evaluate the effect of pH on lysozyme binding. Results showed that maximum adsorption occurred at pH 9, where lysozyme carries a strong positive charge while the membrane is negatively charged, enhancing electrostatic attraction. At this pH, P-BrA demonstrated a lysozyme adsorption capacity of 483.85 mg/g, significantly higher than other tested membranes. Dynamic adsorption studies further confirmed optimal performance under conditions of 10% CEW dilution, pH 9, and a flow rate of 0.1 mL/min. Under these conditions, the purification factor reached 402-fold, with a yield of 91%. Breakthrough curves indicated that the 10% breakthrough point was achieved at 23.3 mL of feed volume, confirming high dynamic capacity.
The scalability of the process was validated using filter holders with different diameters (25 mm and 47 mm).TMEM173 Antibody supplier Consistent purification performance across scales demonstrated the feasibility of large-scale application.PMID:35238624 The membrane also exhibited excellent reusability; after five consecutive cycles of adsorption, elution, and cleaning-in-place (CIP), no significant loss in adsorption capacity was observed. Regeneration with 0.1 M NaOH maintained membrane integrity and functionality.
Elution studies revealed that a two-step salt gradient—0.6 M NaCl followed by 1 M NaCl—achieved near-complete lysozyme recovery (95.43%) with minimal protein contamination. SDS-PAGE analysis confirmed high purity (>98%), and specific activity measurements indicated superior quality compared to commercial standards. These results demonstrate that P-BrA is highly selective for lysozyme, minimizing non-specific binding of other proteins.
In conclusion, the polyacid IEX nanofiber membrane offers a rapid, scalable, and sustainable method for lysozyme purification from CEW. Its high adsorption capacity, excellent selectivity, and reusability make it a competitive alternative to conventional chromatographic techniques. This work highlights the potential of advanced nanomaterials in improving downstream processing in biotechnology applications.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com