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Experimental design of a twin-column countercurrent gradient purification process.

Fabian Steinebach¹, Nicole Ulmer¹, Lara Decker¹

¹Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.

Journal of Chromatography. A

|March 12, 2017

Summary

A new design procedure for multi-column countercurrent solvent gradient purification (MCSGP) chromatography improves both purity and yield for challenging separations. This method efficiently isolates monoclonal antibody charge variants, achieving over 90% purity and yield.

Keywords:

Charge isoforms Continuous chromatography MCSGP Monoclonal antibody

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Area of Science:

Biotechnology
Chemical Engineering
Separation Science

Background:

Batch chromatography processes often face a purity-yield trade-off, limiting efficiency.
Multi-column countercurrent solvent gradient purification (MCSGP) offers a solution to overcome these limitations, especially for complex separations.

Purpose of the Study:

To develop an efficient and reliable design procedure for the twin-column MCSGP process.
To demonstrate the application of this procedure for isolating monoclonal antibody charge variants using ion-exchange chromatography.

Main Methods:

The design procedure is based on a single selected batch chromatogram.
The method focuses on achieving target product recovery from a specific fraction of the batch chromatogram with enhanced yield.
Ion-exchange chromatography was used to isolate the main charge isoform of a monoclonal antibody from Protein A eluate.

Main Results:

The developed MCSGP process achieved over 90% purity and yield for the target monoclonal antibody charge isoform.
Process-related impurities, including host cell proteins (HCP), leached Protein A, and aggregates, were effectively removed.
The study also analyzed the impact of design parameters on purity, yield, productivity, and buffer consumption.

Conclusions:

The developed design procedure provides an efficient method for twin-column MCSGP process implementation.
This approach successfully alleviates the purity-yield trade-off in chromatographic separations.
The findings offer a basis for further process optimization to enhance performance in monoclonal antibody purification.