Modeling Column Regeneration Effects on Dye-Ligand Affinity Chromatography

Abstract

The effect of in-place regeneration of dye-ligand adsorbents on protein adsorption characteristics is presented. Regeneration with chemical treatments and time of exposure determined the protein capacity of the adsorbent, but no effect was observed on its protein binding affinity. Fixed-bed adsorption of bovine serum albumin and its selectivity with respect to lysozyme was studied. Breakthrough curves were measured for protein adsorption on fixed-bed columns and analyzed by a simple model to determine the relevant rate constants for the adsorption process. It was found that forward adsorption rate constant increased exponentially with the chemical treatment exposure time. Column linear gradient elution studies showed that adsorbent selectivity decreased with the chemical treatment exposure time due mainly to column loss of adsorption capacity. The implications of the results on the design and optimization of dye-ligand chromatographic processes are discussed.

Lumped parameter model for prediction of initial breakthrough profiles for the chromatographic capture of antibodies from a complex feedstock

“…However, within the last two decades various mathematical models have been developed describing the breakthrough behaviour of a range of chromatographic systems, such as hydrophobic interaction [1], dye ligand affinity [2,3], ion-exchange [4], and affinity [5,6]. These models can provide an understanding of the equilibrium and mass transfer (MT) characteristics of an adsorbent, and consequently represents powerful tools for targeted optimisation and scale-up.…”

“…Very similar overall mass balances are applied in the published models, and the majority of these employ the Langmuir adsorption isotherm [1][2][3]5,6,8,9] for describing the non-linear favourable kinetics, typically observed with most proteins. Sophisticated models, taking film, pore, or surface diffusion (or various combinations) into account, have been developed [1,5,6,8].…”

“…Most work done so far on development of models describing chromatographic processes has been performed with mono- [3,5,6,8,9] or bi- [10] component systems, and in only one report was a multi-component system analysed, i.e. for the adsorption of lipase from a fermentation broth [1].…”

Protein loading, elution, and resolution behavior in a novel device that integrates ultrafiltration and chromatographic separation

“…Each adsorption experiment was carried out until no appreciable change was observed. This procedure is similar to that described by Chase (1984), Boyer (1991), and Noriega et al (1997).…”

Improved hollow-fibre membranes for dye-affinity chromatography

“…Membrane segments were incubated in 6 M urea in 0.5 M NaOH at room temperature for 6 months according to Noriega et al [15]. Dye leakage was quantified monthly by measuring the absorbance at 610 nm; a molar extinction coefficient of 13.6 for Cibacron Blue F3G-A was assumed.…”