Single-step affinity purification of enzyme biotherapeutics: a platform methodology for accelerated process...

Area of Science:

• Biopharmaceutical Process Development
• Analytical Chemistry
• Biotechnology

Background:

• Downstream purification of non-antibody biologics is a bottleneck, requiring multiple chromatography steps before analysis.
• This multi-step process limits throughput, extends timelines, and increases resource demands.

Purpose of the Study:

• To develop a single-step purification method for enzyme biotherapeutics using immunoaffinity technology.
• To apply predictive analytics for forecasting critical quality attributes (CQAs) of biologics post-purification.
• To demonstrate the platform applicability of the developed methodology.

Main Methods:

• Immunoaffinity purification using polyclonal and monoclonal antibodies as ligands for enzyme biotherapeutics (agalsidase beta and Enzyme 2).
• Isolation of target molecules from cell culture harvest for CQA analysis.
• Development of predictive models based on affinity column data to forecast CQAs after traditional purification.

Main Results:

• Achieved single-step purification of enzyme biotherapeutics to sufficient purity for CQA analysis.
• Successfully applied predictive analytics to forecast CQAs, providing early process development insights.
• Demonstrated successful reapplication of the methodology for a second enzyme biotherapeutic, indicating platform potential.

Conclusions:

• Immunoaffinity purification offers a rapid, high-throughput alternative to multi-step chromatography for enzyme biotherapeutics.
• Predictive analytics integrated with affinity purification can accelerate bioprocess development and decision-making.
• The developed platform supports Quality by Design (QbD) principles and Process Analytical Technology (PAT) integration.