Application of enhanced electronegative multimodal chromatography as the primary capture step for immunoglobulin G...

Area of Science:

• Biotechnology
• Bioseparation
• Protein Purification

Background:

• Direct capture of immunoglobulin G (IgG) from undiluted cell culture fluid is crucial for biopharmaceutical manufacturing.
• Electronegative multimodal chromatography (MCM) with Eshmuno HCX has shown promise for IgG recovery.
• Optimization of chromatography conditions is essential for enhancing purification performance and process efficiency.

Purpose of the Study:

• To systematically investigate the binding properties of Eshmuno HCX to IgG under various pH and salt conditions.
• To enhance the purification performance of IgG capture by optimizing washing parameters.
• To evaluate the overall IgG recovery and purity achieved through the optimized chromatography process.

Main Methods:

• Systematic study of IgG binding to Eshmuno HCX at different pH/salt combinations.
• Optimization of washing pH and conductivity following high-capacity IgG binding.
• Analysis of end-product purity, including non-histone host cell protein (nh-HCP), DNA, and aggregates.

Main Results:

• Purification performance was significantly enhanced by lowering washing pH and conductivity.
• A single polishing step yielded IgG with nh-HCP below 1 ppm and DNA less than 1 ppb.
• The process achieved less than 0.5% aggregates and an overall IgG recovery of 86.2%.

Conclusions:

• The optimized electronegative multimodal chromatography process enables efficient direct IgG capture from cell culture fluid.
• The two-column, non-affinity chromatography process supports direct feed loading without buffer adjustment, boosting productivity.
• This approach offers significant cost-savings and improved overall process efficiency for IgG purification.