Characterization of Aggregation Propensity of a Human Fc-Fusion Protein Therapeutic by Hydrogen/Deuterium Exchange Mass Spectrometry

J Am Soc Mass Spectrom. 2017 May;28(5):795-802. doi: 10.1007/s13361-016-1452-7. Epub 2016 Aug 15.


Aggregation of protein therapeutics has long been a concern across different stages of manufacturing processes in the biopharmaceutical industry. It is often indicative of aberrant protein therapeutic higher-order structure. In this study, the aggregation propensity of a human Fc-fusion protein therapeutic was characterized. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) was applied to examine the conformational dynamics of dimers collected from a bioreactor. HDX-MS data combined with spatial aggregation propensity calculations revealed a potential aggregation interface in the Fc domain. This study provides a general strategy for the characterization of the aggregation propensity of Fc-fusion proteins at the molecular level.Graphical Abstract.

Keywords: Aggregation propensity; Fc fusion protein; Hydrogen/Deuterium exchange mass spectrometry.

MeSH terms

  • Deuterium Exchange Measurement / methods
  • Humans
  • Immunoglobulin Fc Fragments / chemistry*
  • Immunoglobulin G / chemistry*
  • Mass Spectrometry / methods*
  • Models, Molecular
  • Protein Aggregates*
  • Protein Conformation
  • Protein Multimerization
  • Recombinant Fusion Proteins / chemistry


  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Protein Aggregates
  • Recombinant Fusion Proteins