Single-component adsorption of proteins on a cellulose membrane with the phenyl ligand for hydrophobic interaction chromatography

Abstract

Porous structure and adsorption properties of a hydrophobic interaction chromatography adsorbent, Sartobind PhenylTM, were studied using bovine γ-globulin, lysozyme, human immunoglobulin G, ovalbumin, bovine serum albumin, and α-chymotrypsinogen A as tested proteins and ammonium sulphate as a kosmotropic salt promoting hydrophobic interactions. Dynamic experiments carried out in a laboratory membrane module showed that the dynamic binding capacity was independent of the flow velocity in the range of 3–45cm/h but it increased exponentially with the salt concentration in the range of 0.5–1.5M. A micromembrane chromatography module was used to examine the effect of pH on the binding capacity of the hydrophobic membrane. Adsorption equilibria were measured using a static batch method for all proteins but α-chymotrypsinogen A. The Langmuir exponential isotherm was employed to describe the effects of ammonium sulphate and protein concentrations on the adsorbed amount.