Digitally programmable microfluidic valving for autonomous, high-resolution continuous chromatographic purification

Area of Science:

• Biotechnology
• Analytical Chemistry
• Chemical Engineering

Background:

• Continuous microscale purification demands precise fluid handling and temporal control.
• Current chromatography systems offer limited programmability and coordination for purification cycles.

Purpose of the Study:

• To develop a microfluidic continuous protein purification method with digitally programmable valves.
• To enable logic-driven chromatography with precise temporal control and contamination-free fraction discrimination.

Main Methods:

• Utilized digitally programmable inlet (ICV) and collection (CCV) valves for logic-driven chromatography.
• Implemented sub-second buffer switching and deterministic routing across parallel affinity columns.
• Employed temporal gating via CCV for real-time, profile-guided fraction selection.

Main Results:

• Achieved 70-89% purity for GFP-His6 over ten continuous cycles with high stability.
• Demonstrated compatibility with structurally sensitive biologics like His6-tagged TRAIL, preserving functional activity.
• System architecture minimizes dead volume, prevents cross-contamination, and supports diverse chromatographic modes.

Conclusions:

• The developed microfluidic system advances purification platforms toward autonomous, precision-controlled operations.
• This method offers a broadly applicable analytical framework for microscale purification.
• Supports development of next-generation bioseparation and continuous biomanufacturing technologies.