Characterizing Monoclonal Antibody Aggregation Using Charge Detection Mass Spectrometry and Industry Standard Methods.

ABSTRACT: Protein aggregation is a factor in a multitude of neurodegenerative diseases and aggregates of protein-based biotherapeutics can cause toxicity in vivo and adverse patient outcomes. Monoclonal antibody (M)-fluorophore (F) complexes with four different antibody sequences and masses of ∼680 kDa were analyzed using size-exclusion chromatography (SEC) and mass spectrometry using both quadrupole-time-of-flight (QTOF) and charge detection mass spectrometry (CDMS). Higher-order aggregates were not resolved using SEC, but species as large as the MF₂ complex and M₅F₃ were resolved using QTOF and CDMS, respectively. Results from three freeze-thaw cycles and long-term heat stress indicate that both aggregation and degradation occurs. Two of the antibodies form

SUBMITTER: Jordan JS 

PROVIDER: S-EPMC12807537 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature