Demonstration of continuous gradient elution functionality with automated liquid handling systems for high-throughput...

Area of Science:

• Biopharmaceutical Manufacturing
• Process Development
• Chromatography

Background:

• High-throughput systems are crucial for biopharmaceutical process development, enhancing experiment productivity and reducing material use.
• Existing high-throughput purification tools, like RoboColumns on Tecan systems, offer scalability but lack optimized liquid delivery for preparative chromatography.
• Limitations include inability to provide continuous liquid flow and ideal linear gradient conditions, impacting protein chromatography and gradient elution experiments.

Purpose of the Study:

• To describe a modified Tecan Freedom EVO system for high-throughput purification.
• To enable continuous gradient elution capability for RoboColumn experiments.
• To demonstrate the system's ability to generate linear conductivity gradients and provide scalable data.

Main Methods:

• Modification of a Tecan Freedom EVO liquid handling system to provide continuous liquid delivery.
• Application of the modified system to RoboColumn experiments for high-throughput purification.
• Generation of linear conductivity gradients and evaluation of RoboColumn performance against larger formats.

Main Results:

• The modified Tecan system successfully enabled continuous gradient elution for RoboColumn experiments, generating highly linear conductivity gradients.
• RoboColumn performance and product quality data were in agreement with larger, bench-scale chromatography formats for two model purification methods.
• The gradient purification method showed more consistent performance between RoboColumns and larger columns compared to step elution methods.

Conclusions:

• The developed Tecan Freedom EVO system enhances high-throughput purification by enabling continuous gradient elution for RoboColumns.
• This advancement provides scalable and consistent chromatography data, bridging the gap between high-throughput screening and manufacturing scale.
• New insights into discontinuous flow effects on RoboColumn performance were gained, aiding bioprocess development.