Ion exchange chromatography of monoclonal antibodies: Effect of resin ligand density on dynamic binding capacity |
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Authors: | Ann Marie Hardin Chithkala Harinarayan Gunnar Malmquist Andreas Axén Robert van Reis |
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Institution: | 1. Genentech Inc., South San Francisco, CA 94080, USA;2. GE Healthcare Bio-Sciences AB, Björkgatan 30, SE-751 84 Uppsala, Sweden |
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Abstract: | Dynamic binding capacity (DBC) of a monoclonal antibody on agarose based strong cation exchange resins is determined as a function of resin ligand density, apparent pore size of the base matrix, and protein charge. The maximum DBC is found to be unaffected by resin ligand density, apparent pore size, or protein charge within the tested range. The critical conductivity (conductivity at maximum DBC) is seen to vary with ligand density. It is hypothesized that the maximum DBC is determined by the effective size of the proteins and the proximity to which they can approach one another. Once a certain minimum resin ligand density is supplied, additional ligand is not beneficial in terms of resin capacity. Additional ligand can provide flexibility in designing ion exchange resins for a particular application as the critical conductivity could be matched to the feedstock conductivity and it may also affect the selectivity. |
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Keywords: | Ion exchange chromatography Protein Monoclonal antibody Exclusion mechanism Ligand density Pore size Dynamic binding capacity |
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