Purification of monoclonal antibodies (mAbs) expressed in bacteria or mammalian host cells, results in extensive bioburden on the protein A column, in the form of DNA and protein impurities when feed is directly loaded without major pre-treatments. To increase the lifetime of these sensitive protein A resins and to improve the final purity of the target molecule, we have included an upstream multimodal AIEX column. This high-salt-tolerant AIEX provides protection for the protein A capture-step column by removing the majority of bulk impurities before the feed is loaded onto the sensitive resin, thus enhancing the lifetime of this resin. Placing a multimodal AIEX column upstream this column to capture impurities early in the process, bridge the more manual and tedious harvesting and pre-treatment steps with the down-stream purification steps. The majority of hazardous impurities such as host cell DNA/ chromatins, viruses and endotoxins from complex feeds are thus removed.<br/>We will present a case study where we have applied this concept. For the mAb purifications the usual three-step purification was performed but now the AIEX step (usually placed post-protein A) was instead placed upstream the protein A column. This re-placement of the purification steps in the mAb purification scheme is a novel concept called ECP, Enhanced Capture Process.<br/>The case study showed that this new upstream concept is highly efficient giving very pure final eluted target molecules. Furthermore, this concept can also be applied to other applications, such as purification of insulin and other peptides and his-tagged proteins, to prolong the lifetime of resins used in the later purification steps, and also to increase the final purity of the target substance.

Pre-purification of complex feeds using a high-salt tolerant AIEX resin prior to protein A chromatography

ACS Spring 2022

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