Development of operating conditions for protein purification using expanded bed techniques: The effect of the degree of bed expansion on adsorption performance

Abstract

A strategy for the optimization of an expanded bed adsorption process has been developed by studying a model system involving the adsorption of lysozyme onto the adsorbent STREAMLINE SP. The hydrodynamic and adsorption properties of this ion exchange adsorbent in a variety of viscosities of feedstocks have been compared by analyzing bed expansion characteristics, liquid phase dispersion characteristics, equilibrium adsorption isotherms, and mass transfer characteristics. Additionally, the influences of the degree of bed expansion on adsorption performance have been investigated by frontal analysis. In these experiments, viscous feedstocks were simulated bythe inclusion of glycerol in the adsorption buffers. Breakthrough curves for lysozyme were characterized and compared in terms of overall purification processing time and productivity. On the basis of these results, the relative productivities of different operating modes with the same process liquid were found to be almost the same. However, the processing time for each purification cycle decreased with increasing velocity of process liquid. It is demonstrated that an adsorption process carried out at a constant degree of bed expansion (twice its settled bed height, corresponding to bed voidage of 0.7) is more efficient, when characterized by the apparent dynamic binding capacity, than operation at a constant liquid velocity of 300 cm/h. These results have significant implications on the design and operation of the expanded bed adsorption procedures. The advantages and problems encountered in the use of expanded bed techniques for the direct extraction of proteins from unclarified feedstocks are also discussed. 0 1996 John Wiley & Sons, Inc.

Hydrophobic interaction adsorption of hen egg white proteins albumin, conalbumin, and lysozyme

“…The decrease of the protein adsorption was observed on the increase of the protein content in the medium. Chang and Chase [9] and Tong et al [11] described a similar kinetic adsorption behavior for lysozyme in Streamline DEAE resin and BSA in Streamline SP resin, respectively. The reduction in the protein adsorption could be attributed to the elevation of the solution viscosity.…”

“…Adsorption chromatography in batch mode, also called batch adsorption, is a simple process applied to obtain (i) equilibrium and kinetic data of adsorption and desorption processes [9][10][11][12] and (ii) model parameters to be used in the simulation of chromatography process scale-up, such as expanded bed and simulated moving bed techniques. It can be cited, for example, in the intra-particle diffusivity parameter [13].…”

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

“…An early breakthrough was recorded as the linear superficial velocity was increased from 0.5 to 5 cm/min. Chang and Chase [20], and Chase [28] also reported that the shape of breakthrough curves varied with superficial velocity when adsorption kinetics were slow relative to the mass transfer rate. A slower superficial velocity during feedstock loading in a packed bed operation may have assisted in giving more interaction time for proteins and adsorbents, thus led to a higher binding of N protein onto the adsorbent as shown by the results tabulated in Table 3.…”

“…Recovery throughput is another parameter in affinity purification. This refers to the amount of adsorbed protein at 10% breakthrough divided by the volume of adsorbent and the processing time; both were calculated according to Chang and Chase [20]. The time for protein adsorption was recorded once the 10% breakthrough was reached.…”

Large-scale recovery of C-phycocyanin from Spirulina platensis using expanded bed adsorption chromatography

“…This method reduces the amount of protein pretreatment required, and makes the capture of target proteins from cell homogenate feedstock possible [21][22][23], thus achieving a higher product recovery in a shorter time. In the first step, phycocyanins were recovered by the adsorbent while the cell debris, particulates and contaminants in the extract pass through with the upward flow.…”