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Method Separates Oxidized Monoclonal Antibody (MAb) Species
Thermo Fisher Scientific has demonstrated that a hydrophobic interaction chromatography (HIC) column, the Thermo Scientific™ ProPac™ HIC-10 column, is suitable for separating low-concentration species of an intentionally oxidized MAb from the native MAb.
The same method can be used to separate the papain fragments from the intentionally oxidized MAb, as well as the papain fragments of the native MAb. Application Note 1084: Separation of Papain Digests of Oxidized Monoclonal Antibodies details how the ProPac HIC-10 column can be used to resolve antigen-binding fragments (Fab) and nonantigen-binding fragments (Fc) generated by papain digestion, as well as oxidized Fab/Fc from native Fab/Fc.
Many MAbs have been developed as human therapeutics and several others are under development. These MAbs are subject to various post-translational modifications and are susceptible to degradation during storage and delivery. Due to the potential impact of these modifications to MAb safety and efficacy, it is necessary to fully characterize and monitor MAb heterogeneity. Other approaches for the characterization and testing of therapeutic antibodies often are not effective for low-level populations of MAb variants, so this new HIC method delivers a much needed solution that is appropriate for stability studies of therapeutic MAbs.