A Study of the Interaction of HEK-293 Cells with Streamline Chelating Adsorbent in Expanded Bed Operation

Abstract

Expanded bed adsorption (EBA) is an efficient protein purification process reducing time and steps of downstream processing (DSP) since nonclarified culture media can be processed directly without prior treatments such as filtration or centrifugation. However, cells and debris can interact with the adsorbent and affect bed stability as well as purification performance. To optimize EBA operating conditions these biomass/adsorbent interactions have to be understood and characterized. The adsorption of Human Embryonic Kidney cells (HEK 293) on unprimed and nickel-primed metal affinity adsorbent was studied in a closed loop EBA setup. With the unprimed adsorbent, the overall level of interaction observed was nonsignificant. With the nickel-primed adsorbent and an initial cell concentration ranging from 0.08 × 10 6 to 0.2 × 10 6 cells/mL, biomass/adsorbent interaction was found to be moderate and the adsorption apparent first-order kinetic rate constant was determined to be k ) 0.009 to 0.011 min -1 .

Surface energetics to assess biomass attachment onto immobilized metal-ion chromatography adsorbents in expanded beds

“…From a process performance point of view, this could indicate hydrodynamic and sorption performance limitations from example, during expanded bed adsorption of bioproducts. Sorption performance utilizing Chelating/EBA systems has been reported recently [4] and characterized as moderate, in full agreement with this paper and previous work [1].…”

“…biological particles and chromatographic beads are anticipated to play a minor role concerning interaction or agglomeration [2,3]. Nevertheless, evidence of potential sorption performance limitations were already observed for IMAC-EBA systems [4]. Biomass effects during EBA with group-specific adsorbents are poorly understood.…”

Recovery and purification of chitosanase produced by Bacillus cereus using expanded bed adsorption and central composite design

“…We believe that the biomass, resin, and distributor interaction is so strong that it determines the effect on the response variable. The influence of biomass on the hydrodynamic stability of expanded beds has been demonstrated by several studies . The distributor has a strong effect on the flow structure .…”

Growth-dependent surface characteristics of Hansenula Polymorpha: implications for expanded bed adsorption chromatography

“…The effects of the biomassadsorbent interactions of different biomass types and their homogenates on the chromatographic performance of a number of commercial adsorbents (Streamline DEAE, SP, phenyl, and chelating matrices) have been studied extensively [15][16][17][18][19][20]. However, it is crucial to understand these interactions from a fundamental perspective.…”