Combined effect of coulombic and van der Waals interactions in the chromatography of proteins.
We have recently proposed a theoretical framework for the effect of the eluting salt ionic strength of the eluent on the retention factor of proteins in ion-exchange chromatography of proteins. It is based on the solution of the linearized Poisson-Boltzmann equation for two oppositely charged planar surfaces in contact with a salt solution and describes the coulombic interaction between the protein and the oppositely charged stationary-phase surface. At sufficiently high salt concentrations in the mobile phase van der Waals interactions between the protein and the stationary phase become important. In this work we consider the effect of salt on the combined coulombic and van der Waals interactions by combining the electrostatic theory with the theory for van der Waals interactions. The combined theory describes the retention of proteins as a function of eluting salt concentration over a wide salt concentration range. The protein molecules are, according to the proposed theory, held in a diffuse layer close to the stationary phase and are not in a distinct layer, which is assumed in the traditional thermodynamic interpretation of the capacity factor. For this reason, we also examine the thermodynamic interpretation of the capacity factor when it is due to distant dependent interactions.
Ståhlberg
J
J
Astra Pharmaceutical Production AB, Quality Control, Södertälje, Sweden.
Jönsson
B
B
Horváth
C
C
D002852:Chromatography, Ion Exchange; D004563:Electrochemistry; D011506:Proteins / Q000737:chemistry*
D011506:Proteins