A Model for Counterion Binding and Charge Reversal on Protein Surfaces

Jas Kalayan, Richard H. Henchman, Jim Warwicker

Research output: Contribution to journalArticlepeer-review


The structural stability and solubility of proteins in liquid therapeutic formulations is important, especially since new generations of therapeutics are designed for efficacy before consideration of stability. We introduce an electrostatic binding model to measure the net charge of proteins with bound ions in solution. The electrostatic potential on a protein surface is used to separately group together acidic and basic amino acids into patches, which are then iteratively bound with oppositely charged counterions. This model is aimed toward formulation chemists for initial screening of a range of conditions prior to lab-work. Computed results compare well with experimental zeta potential measurements from the literature covering a range of solution conditions. Importantly, the binding model reproduces the charge reversal phenomenon that is observed with polyvalent ion binding to proteins and its dependence on ion charge and concentration. Intriguingly, protein sequence can be used to give similarly good agreement with experiment as protein structure, interpreted as resulting from the close proximity of charged sidechains on a protein surface. Further, application of the model to human proteins suggests that polyanion binding and over-charging, including charge reversal for cationic proteins, is a general feature. These results add to evidence that addition of polyanions to protein formulations could be a general mechanism for modulating solution stability.
Original languageEnglish
Pages (from-to)595-603
JournalMolecular Pharmaceutics
Issue number2
Early online date30 Dec 2019
Publication statusE-pub ahead of print - 30 Dec 2019


  • Zeta potential
  • charge reversal
  • therapeutic protein stability
  • polyvalent ions
  • Electrostatic interactions

Research Beacons, Institutes and Platforms

  • Manchester Institute of Biotechnology


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