Bulk drug substances are frequently stored frozen in stainless steel 316L or Hastelloy® containers.  During the manufacturing process, exposure to ambient temperatures for limited periods of time may be necessary. For the monoclonal antibody (mAb1) used in the present study, extended thermal stress in lab-scale Hastelloy® containers led to a significant increase in high molecular weight species (HMWS, monitored by SEC) and irreducible species (monitored by reduced CE-SDS), which were not representative of the species formed in vial controls or under recommended storage conditions. The nature of the HMWS and irreducible species were further characterized with a suite of analytical assays including peptide mapping, O18-labeled peptide mapping, LC/MS/MS, and IdeS digestion followed by LC/MS analysis. We identified several dityrosine crosslinked sites between the heavy chain Fab and Fc domains in the stressed mAb1 samples. A general increase in oxidation was also observed in heavy chain Fab and Fc domains consistent with metal-catalyzed oxidation. These findings demonstrate that extended thermal stress studies can induce the formation of metal catalyzed oxidation products even when performed in Hastelloy® containers, which have a composition that is significantly lower in iron content than traditional stainless steel 316L.  Therefore, thermal stress studies in metal containers should be undertaken with caution.  <br/>

Identification of dityrosine crosslinking in a monoclonal antibody subjected to thermal stress in lab-scale Hastelloy® containers

254th National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.