A Study of the interaction of HEK-293 cells with streamline chelating adsorbent in expanded bed operation |
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Authors: | Poulin Frédéric Jacquemart Renaud De Crescenzo Gregory Jolicoeur Mario Legros Robert |
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Affiliation: | Biotechnology and pharmaceutical processes research unit (Bio-P2), Department of Chemical Engineering, Ecole Polytechnique, Montréal, Québec, Canada. |
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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 x 10(6) to 0.2 x 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). |
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