Fragmentation of monoclonal antibodies is a frequently observed degradation pathway that can limit the shelf life of recombinant monoclonal antibodies in biopharmaceutical products. We investigate one cause of fragmentation, copper catalysed hydrolysis of a lys222-thr223 bond in the hinge region of an IgG1 mAb. We also developed sample preparation and handling approaches and applied analytical techniques to overcome the solubility limitations of copper in commercially relevant formulations containing phosphate buffer.<br/>Analysis of the kinetics of the degradation pathway over a temperature range of 5-30°C and a copper concentration range of ~50-500 ppb indicated that the fragmentation reaction follows zero-order kinetics and yield products consistent with copper mediated fragmentation. By understanding the fragmentation kinetics, we were able to determine limits on copper impurities that allow for adequate processing time in the drug substance and drug product manufacturing processes while minimizing fragmentation.<br/>Furthermore, we investigate the kinetics of copper removal from the mAb using EDTA as a chelator for copper in solution. By understanding the copper removal kinetics, we were able to determine the necessary contact times and EDTA levels to ensure reduction of mAb bound copper to acceptable levels during drug substance manufacturing.

Copper catalyzed fragmentation of an IgG1 recombinant monoclonal antibody for therapeutic use: using kinetic modeling to fit process to product during technology transfer

255th National Meeting (2018)

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