Evaluation of continuous nonaffinity capture chromatography for a recombinant enzyme-optimization for a changing...

Area of Science:

• Biochemical Engineering
• Downstream Processing
• Bioseparation

Background:

• Recombinant enzyme production often uses perfusion upstream processes, yielding low titers and variable feedstreams.
• Traditional batch chromatography faces challenges with fluctuating enzyme concentrations and impurities.
• Optimizing downstream purification is critical for efficient biomanufacturing.

Purpose of the Study:

• To evaluate the feasibility and economic benefits of continuous capture chromatography for recombinant enzyme purification.
• To develop a design approach for continuous chromatography handling variable feedstreams.
• To statistically characterize and optimize the operating space under constraints.

Main Methods:

• Implemented continuous capture chromatography for downstream processing.
• Utilized process economic modeling and Monte Carlo simulations for cost analysis.
• Compared continuous processing to batch mode and repeated batch column cycling.

Main Results:

• Continuous chromatography demonstrated cost savings compared to batch processing, even with inexpensive ion-exchange resins.
• Economic attractiveness increases for processes with limited column utilization or slow mass transport.
• Continuous processing offers advantages over batch for sharp breakthroughs and short resin lifetimes, though repeated batch cycling also yields savings.

Conclusions:

• Continuous capture chromatography is an economically viable and efficient strategy for downstream processing of recombinant enzymes.
• The approach effectively addresses challenges posed by variable feedstreams common in perfusion upstream processes.
• Continuous processing provides significant advantages when column lifetime is limited or mass transfer is slow.