Orthogonal techniques to study the effect of pH, sucrose and arginine salts on monoclonal antibody physical stability and aggregation during long-term storage

Hristo L. Svilenov*, Alina Kulakova, Matja Zalar, Alexander P. Golovanov, Pernille Harris, Gerhard Winter

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Understanding the effects of additives on therapeutic protein stability is of paramount importance for obtaining stable formulations. In this work, we apply several high- and medium-throughput methods to study the physical stability of a model monoclonal antibody at pH 5.0 and 6.5 in the presence of sucrose, arginine hydrochloride and arginine glutamate. In low ionic strength buffer, the addition of salts reduces the antibody colloidal and thermal stability, attributed to screening of electrostatic interactions. The presence of glutamate ion in the arginine salt partially reduces the damaging effect of ionic strength increase. The addition of 280 mM sucrose shifts the thermal protein unfolding to a higher temperature. Arginine salts in the used concentration reduce the relative monomer yield after refolding from urea, while sucrose has a favorable effect on antibody refolding. In addition, we show 12-month long-term stability data and observe correlations between thermal protein stability, relative monomer yield after refolding and monomer loss during storage. The monomer loss during storage is related to protein aggregation and formation of subvisible particles in some of the formulations. This study shows that the effect of commonly used additives on the long-term antibody physical stability can be predicted using orthogonal biophysical measurements.
Original languageEnglish
JournalJournal of Pharmaceutical Sciences
Volume109
Issue number1
Pages (from-to)584-594
ISSN0022-3549
DOIs
Publication statusPublished - 2020

Keywords

  • Monoclonal antibody(s)
  • pH
  • Sucrose
  • Arginine
  • Physical stability
  • Protein aggregation
  • Protein formulation
  • Fluorescence spectroscopy
  • Light scattering (dynamic)
  • Nuclear magnetic resonance (NMR) spectroscopy

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