title: Modeling of preparative reversed-phase HPLC of insulin.
abstract: The adsorption isotherms of three recombinant proteins, human insulin, porcine insulin, and Lispro, were measured by frontal analysis on a YMC-ODS C18 column with an aqueous solution at 31% acetonitrile (0.1% TFA) as the mobile phase. The retention behavior of insulin, its related molecular structure, its conformation, and its aggregation in this phase system are discussed. The experimental isotherm data were fitted to the Langmuir, the Langmuir-Freundlich, and the Toth models. The results allow for a quantitative comparison of the saturation capacities, the equilibrium constants, and the exponents that represent the heterogeneity of the stationary phase obtained for the different insulin variants studied. The Toth model provided the best fit of the experimental data. The overloaded band profiles were calculated using the lumped pore diffusion and the equilibrium-dispersive model of chromatography. An excellent agreement between calculated and experimental profiles was demonstrated.
authors.affiliation: Department of Chemistry, The University of Tennessee, Knoxville, Tennessee 37996-1600, USA.
authors.forename: Xiaoda
authors.initials: X
authors.lastname: Liu
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Kaczmarski
authors.forename: Alberto
authors.initials: A
authors.lastname: Cavazzini
authors.forename: Pawel
authors.initials: P
authors.lastname: Szabelski
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zhou
authors.forename: Georges
authors.initials: G
authors.lastname: Guiochon
mesh_terms: D000327:Adsorption; D000818:Animals; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004058:Diffusion; D006801:Humans; D007328:Insulin / Q000737:chemistry* / Q000302:isolation & purification*; D007700:Kinetics; D016015:Logistic Models; D008956:Models, Chemical; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D013552:Swine; D013696:Temperature
chemical_list: D007328:Insulin; D011994:Recombinant Proteins