Water distribution and clustering on lyophilized IgG1 surface: insight from molecular dynamics simulations

Shaoxin Feng, Günther H.J. Peters, Satoshi Ohtake, Christian Schoneich, Evgenyi Shalaev*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review


Water has a critical role in the stability of the higher-order structure of proteins. In addition, it is considered to be a major destabilization factor for the physical and chemical stability of freeze-dried proteins and peptides. Physical chemical aspects of protein/water relationships are commonly studied with the use of water vapor sorption isotherms for amorphous lyophilized proteins, which, in turn, are commonly analyzed using the Brunauer-Emmett-Teller (BET) equation to obtain the parameters, Wm and CB. The parameter Wm is generally referred to as the "monolayer-limit of adsorption" and has a narrow range of 6-8% for most proteins. In this study, the water distribution on an IgG1 surface is investigated by molecular dynamics (MD) simulations at different water contents. The monolayer of water molecules was found to have limited coverage of the protein surface, and the true monolayer coverage of the protein globule actually occurs at a hydration level above 30%. The distribution of water molecules on the IgG1 surface is also highly heterogeneous, and the heterogeneity is not considered in the BET theory. In this study, a mechanistic model has been developed to describe the water vapor sorption isotherm. This model is based on the analysis of the hydrogen bonding network extracted from the MD simulations. The model is consistent with the experimental Type-II isotherm, which is usually observed for proteins. The physical meaning of the BET monolayer was re-defined as onset of water cluster formation. A simple model to calculate the onset water level, Wm, is proposed based on the hydration of different amino acids, as determined from the MD simulations.
Original languageEnglish
JournalMolecular Pharmaceutics
Issue number3
Pages (from-to)900-908
Publication statusPublished - 2020


  • Water sorption
  • Lyophilized protein
  • Molecular dynamic simulation
  • Water distributions on protein surface
  • BET monolayer

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