Examination of protein adsorption in fluidized bed and packed bed columns at different temperatures using frontal chromatographic methods

Abstract

The influences of various experimental parameters on the dynamic adsorption capacity (DAC) and the dynamic adsorption rate (DAR) of a biomimetic affinity silica‐based adsorbent in fluidized and packed bed columns operated under plug flow conditions and at different temperatures have been investigated with different inlet concentrations of hen egg white lysozyme (HEWL) and human serum albumin (HSA). The DACs as well as the DARs of both the fluidized and packed beds were examined at 10% saturation (i.e., at the QB value) and the experimental data compared with the corresponding data obtained from batch equilibrium adsorption procedures. Parameters examined included the fluid superficial velocity and protein concentration and their effect on the binding capacity and column efficiency. Consistent with various results reported from this and other laboratories on the behavior of biospecific affinity adsorbents derived from porous silica and zirconia particles, adsorbents prepared from Fractosil 1000 were found to exhibit appropriate rheological characteristics in fluidized bed systems under the experimental conditions. Moreover, changes in temperature resulted in a more significant effect on the breakthrough profiles of HSA compared to HEWL with the immobilized Cibacron Blue F3G‐A with Fractosil 1000 adsorbent. This result suggests that temperature effects can possibly be employed profitably in some processes as part of a strategy to enhance column performance with fluidized bed systems for selective recovery of target proteins. At relatively low superficial velocities of the feed, the DARs with HEWL and HSA were similar for both the fluidized and packed bed column systems, whereas, at high superficial velocities, the DARs for these proteins were larger with the packed bed columns. © 1998 John Wiley &, Inc. Biotechnol Bioeng 58:35–46, 1988.

Purification of histidine-tagged nucleocapsid protein of Nipah virus using immobilized metal affinity chromatography

“…However, higher amount of N protein was eluted with the buffer containing 300 mM imidazole (lanes [12][13][14][15][16], and the thickest band was observed on lane 13 (Fig. 1).…”

“…The protein adsorption on IMAC is governed by various factors such as the type of chelating ligand, metal ion, the surrounding chemical environment and experimental parameters on the dynamic binding capacity [11][12][13][14]. Furthermore, Sharma et al [12] demonstrated that the role of ionic strength and pH of the chromatographic medium is significant in governing the binding interaction between the protein and the adsorbent.…”

“…The performance of a chromatography process such as dynamic binding capacity and recovery throughput can be assessed by frontal breakthrough analysis. Finette et al [14] examined frontal breakthrough measurements in packed bed IMAC with different flow rates and different inlet concentrations of hen egg white lysozyme and human serum albumin.…”

Evaluation of commercial chromatographic adsorbents for the direct capture of polyclonal rabbit antibodies from clarified antiserum

“…In experiments with 50% (v/v) antiserum, the significant drop in MabSelect's Ig binding capacity from 48 to 37 mg mL −1 , that was observed on shifting from ambient to 4-8 • C (Fig. 3), is most likely attributable to reduced mass transfer [34,35] at low temperature combined with an increased tendency for support fouling to occur. It is plausible that the latter effect could, at least in part, account for: (i) MabSelect's further reduction in binding capacity to just 22 mg mL −1 on raising the antiserum strength two-fold (i.e.…”

Simulation of the breakthrough curves for the adsorption of α-lactalbumin and β-lactoglobulin to SP Sepharose FF cation-exchanger

“…either the rate of adsorbate-adsorbent interaction or the rate of adsorbate diffusion (pore or film) as controlling the overall adsorption mechanism [1][2][3][4][5]. Arve and Liapis [6] developed a sophisticated model for biospecific affinity chromatography considering three rate controlling mechanisms simultaneously, two for film and porediffusion mass transfer mechanisms and one for the interaction between adsorbate and adsorbent.…”