A Multi-Method Approach to Assess the Self-Interaction Behavior of Infliximab

J Pharm Sci. 2021 May;110(5):1979-1988. doi: 10.1016/j.xphs.2021.02.002. Epub 2021 Feb 5.


Attractive self-interaction processes in antibody formulations increase the risk of aggregation and extraordinarily elevated viscosity at high protein concentrations. These challenges affect manufacturing and application. This study aimed to understand the self-interaction process of Infliximab as a model system with pronounced attractive self-interaction. The association mechanism was studied by a multi-method approach comprising analytical ultracentrifugation, dynamic light scattering, small angle X-ray scattering, self-interaction bio-layer interferometry and hydrogen-deuterium exchange mass spectrometry. Based on our results, both Fab and Fc regions of Infliximab are involved in self-interaction. We hypothesize a mechanism based on electrostatic interactions of polar and charged residues within the identified areas of the heavy and the light chain of the mAb. The combination of fast and reliable screening methods and low throughput but high resolution methods can contribute to detailed characterization and deeper understanding of specific self-interaction processes.

Keywords: AUC; BLI; HDX-MS; Infliximab; SAXS; Self-association; Self-interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies*
  • Dynamic Light Scattering
  • Infliximab
  • Ultracentrifugation
  • Viscosity


  • Antibodies
  • Infliximab