The purification of lysozyme from chicken egg white (CEW) remains a critical challenge in bioprocessing due to the complexity of the feedstock and the need for high purity and yield. This study evaluates the performance of a bromoacetic acid-functionalized polyacid ion exchange nanofiber membrane (P-BrA) in a scalable membrane chromatography system, focusing on process consistency across different operational scales. The investigation was conducted using Pall filter holders with inner diameters of 25 mm and 47 mm, enabling direct comparison of adsorption capacity, recovery efficiency, and purification factor under identical conditions.
The P-BrA membrane was prepared by electrospinning PAN fibers followed by sequential modification with ethylene diamine and bromoacetic acid. Scanning electron microscopy confirmed a uniform fibrous morphology with an average diameter of 467.3 nm. FTIR analysis verified successful functionalization, showing characteristic peaks corresponding to amide and carboxyl groups. The membrane exhibited a porosity of 83.67% and a mean pore size of 457.1 nm, facilitating rapid convective flow and efficient protein transport. Thermogravimetric analysis revealed that the membrane remained stable at temperatures below 320°C, ensuring reliability during processing.
Batch experiments established optimal conditions for lysozyme adsorption: pH 9, 10% CEW dilution, and 0.1 mL/min flow rate. Under these parameters, the dynamic adsorption capacity reached 4.41 × 10⁶ U/g at the 10% breakthrough point. The purification factor was 402-fold, and the yield was 91%. When scaled up to the 47 mm filter holder, the effective membrane area increased from 3.7 cm² to 9.6 cm², allowing a loading volume of 26 mL.VEGF-A Antibody Technical Information Despite the larger scale, the purification profile remained consistent—lysozyme breakthrough was only 0.RIP3 Antibody Autophagy 56%, and total protein loss was minimal at 83.PMID:35112351 09%.
The two-step elution protocol—using 0.6 M NaCl followed by 1 M NaCl—achieved a recovery yield of 89.60% and a purification factor of 362.90 for the 47 mm system, demonstrating excellent scalability. SDS-PAGE analysis confirmed high purity, with a single dominant band corresponding to lysozyme. The specific activity of the purified product reached 9.12 × 10⁵ U/mg, exceeding that of commercial standards.
Membrane reusability was assessed over five cycles. After each cycle, regeneration with 0.1 M NaOH and CIP treatment restored full functionality. Adsorption capacity remained unchanged within ±3%, confirming robustness and long-term stability. The permeate volume flux (JP,V) was maintained at 0.027 mL/cm²·min throughout all runs, and the volume flux recovery ratio (FRR,V) was consistently near 100%, indicating no significant fouling or pore blockage.
Pressure drop across the membrane remained below 0.01 MPa, even at higher flow rates, highlighting the low resistance inherent to the nanofiber structure. The absence of clogging or biofouling was attributed to the open, interconnected pore network and hydrophilic surface properties. These results confirm that the P-BrA membrane maintains its performance across multiple scales without compromising selectivity or throughput.
In summary, this study demonstrates that the P-BrA nanofiber membrane enables highly efficient, scalable lysozyme purification from CEW. The process is not only consistent across different reactor sizes but also exhibits excellent reusability and operational stability. These findings support the industrial viability of membrane chromatography using functionalized nanofibers as a next-generation platform for protein purification.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
