Capillary electrophoresis for automated on-line monitoring of suspension cultures: Correlating cell density, nutrients and metabolites in near real-time

• Automated SI-CE system for monitoring suspension cultures every 30 min over a 4 day period.
• CE-C4D separation method using 60 mM TEA/10 mM CAPS, pH 12.4 in HDMB/PSS/HDMB coated PMMA capillary stable for at least 24 h.
• CE-C4D separation using 60 mM TEA/10 mM CAPS, pH 12.4 in HDMB/PSS/HDMB coated PMMA capillary stable for at least 24 h.
• Microfluidic devices enable digital cell density measurement and extraction of a cell-free sample using an H-filter.
• Cell density data provide valuable insight in the metabolic state of the cells.

Increasingly stringent demands on the production of biopharmaceuticals demand monitoring of process parameters that impact on their quality. We developed an automated platform for on-line, near real-time monitoring of suspension cultures by integrating microfluidic components for cell counting and filtration with a high-resolution separation technique. This enabled the correlation of the growth of a human lymphocyte cell line with changes in the essential metabolic markers, glucose, glutamine, leucine/isoleucine and lactate, determined by Sequential Injection-Capillary Electrophoresis (SI-CE). Using 8.1 mL of media (41 μL per run), the metabolic status and cell density were recorded every 30 min over 4 days. The presented platform is flexible, simple and automated and allows for fast, robust and sensitive analysis with low sample consumption and high sample throughput. It is compatible with up- and out-scaling, and as such provides a promising new solution to meet the future demands in process monitoring in the biopharmaceutical industry.

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