sepharose and tungsten-carbide

Expanded bed adsorption (EBA) is a promising technology to capture proteins directly from unclarified feedstock. In order to better understand liquid mixing along the bed height in expanded beds, an in-bed sampling method was used to measure residence time distribution at different bed heights. A 2cm diameter nozzle column was tested with agarose raw beads (3% crosslinked agarose containing tungsten carbide). Two settled bed heights (11.5 and 23.1cm) with different expansion factors (1.4-2.6) were investigated and the number of theoretical plates (N), the height equivalent of theoretical plate (HETP) and the local effective axial dispersion coefficient (Dax) were calculated for each bed height-defined zone. The effects of expansion factor, settled bed height and mobile phase were evaluated. The results showed that N increased with the increase of expansion factors, but Dax was unaffected under fixed bed heights. Dax and HETP were found similar as a function of relative bed height for two settled bed heights tested. Higher mobile phase viscosity resulted in stronger axial dispersion. In addition, the local effective Dax under the expansion factor near 2.0 had a different profile which showed minimum values at 0.6-0.8 relative bed height, and the potential mechanism was discussed. These results would be useful for the characterization of axial dispersion and modeling protein adsorption in expanded beds under varying operation conditions.

Topics: Adsorption; Chromatography, Liquid; Sepharose; Tungsten Compounds; Viscosity

Expanded bed adsorption (EBA) is an innovative chromatography technology that allows the adsorption of target proteins directly from unclarified feedstock, and the most important property of an expanded bed is the perfectly classified fluidization of resin beads in the column. Due to the variation of both size and density of bulk resin beads, the axial distributions of bead size, bead density and bed voidage are the inherent characteristics of an expanded bed. However, the understanding on these properties is quite limited. In this study, raw beads (3% crosslinked agarose containing tungsten carbide) and 2cm-diameter nozzle column were used as the model system and mean bead size, bead density and local bed voidage along the bed height were measured systematically with the in-bed sampling method for two settled bed heights (11.5 and 23.1cm) and different expansion factors (1.4-2.6). With the increase of bed height, mean bead size and wet density of the beads decreased from 140 to 90μm and from 4 to 2g/ml, respectively. The local bed voidage increased from 0.6 to 0.9 with the increasing bed height. The relative bed height and relative bed voidage were introduced to describe the general rule of axial distribution. Some empirical equations were used to correlate the mean bead size, bead density and local bed voidage along the bed height with the standard deviations of 10.6%, 6.1% and 5.5, respectively. In addition, a general equation was proposed to predict the axial distributions of bead size, bead density and local bed voidage in the column with standard deviations less than 10% for most of the experimental data, which would be useful for the characterization of resin beads distribution in an expanded bed under varying operation conditions.

Topics: Adsorption; Algorithms; Chromatography; Equipment Design; Particle Size; Porosity; Sepharose; Tungsten Compounds