Protein refolding at high concentration usually leads to formation of protein precipitates. Direct filtration of these refold materials is often very challenging, namely low filter capacity and high filtration variability. The use of EBA here eliminated the need to filter the original refold materials. Instead, filtration was performed only on elution pools with substantially reduced volume (>3-fold) and turbidity (>30-fold), hence much easier filtration. The unique property of the resin used (FastLine SP IEX) allowed EBA to be operated at very high linear velocity (800-1600 cm/hr), while still providing sufficient selectivity and impurity clearance. Furthermore, the relatively broad elution profile of EBA is particularly suitable for such proteins of low solubility as the one studied here that readily precipitated in packed bed due to sharp elution profile. This work demonstrates that the use of EBA helps debottleneck downstream productivity by achieving efficient clarification and high-capacity capture in a single step.

Clarification and capture of high concentration refold pools for E.coli based therapeutics using expanded bed adsorption chromatography

243rd National Meeting (2012)

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