HIGH EFFICIENCY AFFINITY CAPTURE AND PURIFICATION OF IMMUNOGLOBULIN G USING MAGNETIC PARTICLES WITH BIOINSPIRIED PEPTIDE LIGANDS
Embargo until
2023-05-01
Date
2019-01-23
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Publisher
Johns Hopkins University
Abstract
While the upstream production rate of monoclonal antibodies has been steadily increasing over time, downstream processing has become a bottleneck in industrial antibody production even with improvements made in recent years to decrease production costs and increase processing capacities. In this regard, magnetic separations have the potential to complement traditional chromatography methods. We designed magnetic beads (MBs) functionalized with Z33 ligands (MB-Z33), for Immunoglobulin G separation and purification. Through Fourier-Transform Infrared Spectroscopy (FTIR), dynamic light scattering (DLs), zeta potential and TEM imaging, we confirmed the success of functionalization. A theoretical capacity (Qmax), for human IgG1 was demonstrated to be 442 ± 16 mg IgG/g beads and a high binding affinity (Ka) of 1.59 x 106 M-1 was achieved. Reutilization and recycling of the beads further showed consistently high yields with over 89% after 6 cycles. FITC-IgG was then used for binding visualization to confirm the low nonspecific binding of bare MBs and the ability for MB-Z33 to bind and elute IgG. Lastly, the operational feasibility of purifying IgG in clarified CHO cell culture harvest bulk was demonstrated with high IgG recovery yield and selectivity.
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Keywords
Magnetic particles, peptide ligands, IgG-binding, purification