High-throughput screening of chromatographic separations: I. Method development and column modeling.

Area of Science:

• Biopharmaceutical Manufacturing
• Protein Purification
• Chromatography

Background:

• Protein biopharmaceutical purification is complex, facing challenges from tight timelines and extensive experimental needs.
• Traditional chromatography method development requires numerous column runs, hindering efficiency.

Purpose of the Study:

• To develop and evaluate an efficient, high-throughput batch-binding method for screening chromatographic purification conditions.
• To optimize parameters for a 96-well format screening technique using robotic liquid handling.

Main Methods:

• A robotic liquid-handling system was used to dispense chromatographic resins into filter plates for batch binding.
• Key factors evaluated included resin volume reproducibility, mixing contact time, and liquid carry-over after centrifugation.
• The method was optimized for resin volume (50 or 100 microL), incubation time (20 min), and hold-up volume assessment.

Main Results:

• The optimized batch-binding method demonstrated high-throughput screening capabilities.
• Performance was comparable to traditional chromatographic column separation for cGMP protein purification.
• Factors like resin volume, contact time, and hold-up volume were quantified and optimized.

Conclusions:

• The developed high-throughput batch-binding method offers an efficient alternative for screening protein purification conditions.
• This technique can be integrated with kinetic and thermodynamic studies for enhanced chromatographic process development.
• The method supports faster and more effective optimization of biopharmaceutical purification processes.