On the solubility of proteins as a function of pH: Mathematical development and application

Thermodynamic relations between the solubility of a protein and the solution pH are presented in this work. The hypotheses behind the development are that the protein chemical potential in liquid phase can be described by Henry's law and that the solid–liquid equilibrium is established only between neutral molecules. The mathematical development results in an analytical expression of the solubility curve, as a function of the ionization equilibrium constants, the pH and the solubility at the isoelectric point. It is shown that the same equation can be obtained either by directly calculating the fraction of neutral protein molecules or by integrating the curve of the protein average charge. The methodology was successfully applied to the description of the solubility of porcine insulin as a function of pH at three different temperatures and of bovine β-lactoglobulin at four different ionic strengths.

► An expression for the solubility of proteins as a function of pH is developed.
► The solubility is calculated with information of the primary protein structure.
► The methodology was applied to the calculation of solubility of two proteins.
► Differences are mostly within experimental uncertainty.
► The developed expressions are exact provided Henry's law is valid.