Protein separation in carousel multicolumn setup. Performance analysis and experimental validation

Area of Science:

• Biochemical Engineering
• Chromatographic Separations
• Process Optimization

Background:

• Batch chromatography faces limitations in periodic protein separations.
• Carousel Multi-Column Setup (CMS) was previously proposed to mimic countercurrent flow.
• Synchronized fluxes in CMS ensure continuous feed but reduce the operating window.

Purpose of the Study:

• To improve the performance of the Carousel Multi-Column Setup (CMS).
• To investigate the impact of flow rate gradients and feed concentration on CMS performance.
• To develop a practical procedure for protein separation using CMS.

Main Methods:

• Theoretical analysis and experimental validation of CMS.
• Systematic variation of flow rate gradient and feed concentration.
• Case study: separation of a ternary protein mixture (cytochrome C, lysozyme, IgG) using hydrophobic interaction chromatography.

Main Results:

• Altering flow rate gradient and feed concentration improved separation selectivity and extended the operating window.
• Feed concentration was identified as the most influential parameter for process performance.
• A practical protocol for protein separation in CMS was successfully developed.

Conclusions:

• The Carousel Multi-Column Setup (CMS) offers a viable approach for continuous protein separation.
• Optimizing feed concentration is crucial for enhancing selectivity and operational efficiency in CMS.
• The developed procedure facilitates the practical implementation of CMS for protein purification.