The microbial production of biotherapeutics represents a critical part of the rapidly evolving biotechnology industry. Around 30% of today’s biopharmaceuticals are expressed in the gram-negative bacterium <i>E. coli</i>. The primary strategy of intracellular protein expression in bacteria requires cell disruption for product release. This complicates downstream processing due to a massive release of process related impurities. While some classes of impurities, such as host cell proteins (HCPs) can be effectively removed by conventional chromatography, other contaminants, such as DNA and endotoxin present a more challenging task due to their physical and chemical characteristics. Yet, being able to significantly decrease their concentration early in the process can make the purification process simpler, more robust, and more inexpensive.<br/>The present study introduces a new strategy of removing contaminants very soon in the process by using a chromatographic clarification approach. Meaning by using a non-woven material having anion exchange characteristics, it is possible to capture soluble and insoluble contaminants based on their charge as well as their size. This allows purification and clarification at the same time, thus reducing the key process related contaminants already at the clarification stage in the production process. We report a DNA clearance of more than 2-log reduction, a HCP removal of at least 0.2-logs and an endotoxin decrease of about 7-logs.<br/>To summarize the present work shows that this strategy is both scalable and platformable, as it utilizes commercial depth filtration and chromatographic clarification technologies to facilitate early impurity reduction.

Chromatographic clarification: Early impurity removal in recombinant protein production

2019 Spring National Meeting

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