349622 Studies on Increasing the Binding Capacity of a Peptide Affinity Absorbent for Antibody Purification

Monday, November 4, 2013
Grand Ballroom B (Hilton)
Tracy Hendrick1, Ruben Carbonell1 and Amith D. Naik2, (1)Chemical Engineering, North Carolina State University, Raleigh, NC, (2)Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC

Antibodies have emerged as the most important class of biopharmaceuticals for effective treatment of different diseases such as cancer and rheumatoid arthritis. Biomanufacturing of an antibody, immunoglobulin G (IgG), based drugs involves purification using Protein A chromatographic resins, which are expensive and chemically labile. Our laboratory had earlier identified a robust linear hexapeptide having specificity for the antibody. This study focuses on enhancing the binding capacity of the peptide ligand affinity adsorbent. In addition to the peptide ligand, there are several other factors affecting antibody purification such as the nature of the base resin, pore size of the resin, spacer arms, and the coupling chemistry. The work presented explores how the binding capacity of IgG is affected by the nature of the resin, the distance between the resin and ligand, and the activation chemistry. Commercial agarose based resins with amine functional groups already bound to the resin were found to have a low binding capacity so experiments were performed on hydroxyl-based resins. This allowed exploration of how different spacer arms affected the binding capacity. Through these experiments, it was determined that one proprietary spacer arm had the highest binding capacity. The current investigation is attempting to optimize the amount of linker added to the resins by adding various molar excesses of ligand relative to the activated groups on the resins and seeing the effects on the IgG binding capacity.

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