Enzyme purification requires multiple chromatographic steps to achieve high purity. These multi-step purifications lead to significant resource requirements during process development and often affect process understanding and design space. While Protein A allows for single-step isolation of Mabs, no such affinity resin is commercially available for non-Mabs, such as a recombinant enzyme, that may not share common features like antibodies. To address this, an affinity methodology was developed to achieve analytical scale single-step purification of two recombinant enzymes. Affinity column development was optimized to maximize process efficiency. Product quality analyses demonstrated that affinity purified samples were predictive of CQAs for drug substance produced using the multi-step purification train. The ability to purify these enzymes with a single-step process significantly reduces FTE and resource requirements. Potential applications of the analytical affinity column include upstream and downstream process development, process understanding improvements, implementation in high throughput platforms, and real-time bioreactor monitoring (PAT).

Affinity-based analytical purification of enzyme biotherapeutics: Methodology and process development applications

245th National Meeting (2013)

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