Comparative analysis of the methods used for finding surface energy to investigate protein interaction behavior on chromatographic supports

Abstract

Hydrophobic interaction chromatography, an important and effective purification strategy, is generally used for the purification of variety of biomolecules. A basic understanding of the protein interaction behavior is required to effectively separate these biomolecules. A colloidal type extended Derjaguin, Landau, Verwey, and Overbeek calculations were utilized to study the interactions behavior of model proteins to commercially available hydrophobic chromatographic materials that is, Toyopearl Phenyl 650C and Toyopearl Butyl 650C. Physicochemical properties of selected model proteins were achieved by contact angle and zeta potential measurements. The contact angle of chromatographic materials used was achieved through sessile drop method on disrupted beads and capillary penetration method (CPM) on intact beads. The surface properties were further used to calculate the interactions of the proteins to chromatographic supports. The calculated secondary energy minimum of the proteins with the chromatographic materials (from the contact angle values determined through both methods can be correlated with the retention volumes from the real chromatography. The secondary energy minimum values are higher for each protein to the chromatographic materials calculated from the inputs derived through sessile drop method compared to CPM. For instance, immunoglobulin G has secondary energy minimum value of 0.17 kT compared to 0.11 kT, obtained through sessile drop method and CPM, respectively. Average relative values of the energy minimum calculated for all proteins are as 1.51 kT and 1.29 kT for Toyopearl Butyl 650C and Toyopearl Phenyl 650C, respectively, as a conversion factor for estimation of secondary energy minimum for both methods.

Understanding the interaction of proteins to ion exchange chromatographic supports: A surface energetics approach

“…A recent study has also been published on the comparison of different surface energy determination methods. An interconversion factor was proposed to convert the secondary energy minimum obtained through SDM and CPM 20 …”

“…Adsorption of colloidal particles (proteins, biomass) depends on the surface properties of both biological molecules and chromatographic materials 23,24 . Different components of surface energy can be calculated from the experimentally determined contact angle (for the determination of Lifshitz van der Waals and acid base components of surface energy) and zeta potential values (for the determination of electrostatic component of surface energy) 2,20,25 . Various studies reported that the adsorption of biomolecules onto ion exchange chromatographic materials is mainly influenced by the zeta potential of the biomolecules and adsorbents 26,27 .…”

“…Similarly, the said approach has been used to explain the interaction of proteins to monoliths 21 . A recent study is published that explains a comparison of different methods used for the determination of surface energies of colloidal particles 20 . This theory has also been used to study bovine serum interaction to different solid surfaces 33 .…”

“…15 Several other attempts have been reported to reduce the associated challenges. Thermodynamics approach, 16 Sogami-Ise, 17 adsorption, 18 DLVO and XDLVO (extended Derjaguin, Landau, Verwey, and Overbeek) approach [19][20][21] are few among the applied strategies.…”

“…23,24 Different components of surface energy can be calculated from the experimentally determined contact angle (for the determination of Lifshitz van der Waals and acid base components of surface energy) and zeta potential values (for the determination of electrostatic component of surface energy). 2,20,25 Various studies reported that the adsorption of biomolecules onto ion exchange chromatographic materials is mainly influenced by the zeta potential of the biomolecules and adsorbents. 26,27 Similarly, some more reports published that highlight the importance of zeta potential in understanding the interaction behavior by characterizing the stationary phase with zeta potential.…”