Examination of protein adsorption in fluidized bed and packed bed columns at different temperatures using frontal chromatographic methods.
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.
Finette
G M
GM
Centre for Bioprocess Technology, Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, 3168 Victoria, Australia.
Mao
Q M
QM
Hearn
M T
MT
D000327:Adsorption; D000818:Animals; D002645:Chickens; D002845:Chromatography / Q000379:methods*; D006801:Humans; D009113:Muramidase / Q000302:isolation & purification; D011485:Protein Binding; D012709:Serum Albumin / Q000302:isolation & purification; D013696:Temperature
D012709:Serum Albumin; D009113:Muramidase