An adaptation of hydrophobic interaction chromatography for estimation of protein solubility optima.
This study describes an adaptation of hydrophobic interaction chromatography (HIC) that can be used to estimate protein solubility optima. The method does not support determination of absolute, e.g. quantitative solubility, however it does provide a basis for identifying the salt concentration, pH, or additive concentrations that support the highest relative solubility. The magnitude of a given salt's effects are consistent with its ranking in the Hofmeister series. IgG in solutions of strong 'precipitating' salts exhibits a classical salting-in/salting-out curve, described by a solubility minimum at low ionic strength, increasing to a well-defined maximum, and then losing solubility with further elevation of salt concentration. The direct effect of pH on protein solubility, as well as its indirect effect via modification of the ionic equilibria of dissolved salts, can also be tracked. Cooperative effects of solubility modifiers such as amino acids can likewise be assessed. The technique can be a useful tool in the development of liquid formulations for protein pharmaceuticals.
Gagnon
P
P
Validated Biosystems Incorporated, Tucson, AZ 85750, USA. pete@validated.com
Mayes
T
T
Danielsson
A
A
D002626:Chemistry, Pharmaceutical; D002845:Chromatography / Q000379:methods*; D019793:Fluorescein; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry; D019999:Pharmaceutical Solutions; D011506:Proteins / Q000737:chemistry*; D012492:Salts; D012995:Solubility; D013050:Spectrometry, Fluorescence
D007074:Immunoglobulin G; D019999:Pharmaceutical Solutions; D011506:Proteins; D012492:Salts; D019793:Fluorescein