High-Throughput Extracellular Vesicle Isolation Using Plate-Based Size Exclusion Chromatography and Automation

Area of Science:

• Biotechnology
• Nanomedicine
• Biochemistry

Background:

• Extracellular vesicles (EVs) are promising biomarkers found in biofluids like plasma.
• Current EV isolation methods, such as size exclusion chromatography (SEC), lack the throughput for clinical applications.
• Reproducible and scalable EV isolation is crucial for biomarker discovery and diagnostics.

Purpose of the Study:

• To adapt SEC into a plate-based format for increased throughput.
• To develop an automated platform for reproducible EV isolation from clinical samples.
• To enable high-throughput analysis of EV biomarkers.

Main Methods:

• Adapted SEC to a 24-well plate format using resin-packed frits.
• Optimized EV separation from free proteins (e.g., Albumin) by measuring EV markers (CD63, CD81).
• Automated the plate-based SEC process using liquid handling platforms.

Main Results:

• Demonstrated successful separation of EVs from free proteins in the 24-well plate format.
• Achieved high reproducibility with the automated platform.
• Successfully applied the automated platform to analyze tetraspanins CD63 and CD81 in plasma samples.

Conclusions:

• The developed automated, plate-based SEC method provides a scalable and reproducible solution for EV isolation.
• This platform significantly increases throughput, enabling hundreds of samples per day.
• The method facilitates EV biomarker discovery and diagnostic applications in clinical settings.