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Adeno-associated viral vector purification using a centrifugal microfluidic system: towards workflow automation for

Matthias Geissler^1,2, Lidija Malic^1,2, Liviu Clime^1,2

¹Medical Devices Research Centre, National Research Council of Canada, 75 de Mortagne Boulevard, Boucherville, QC, J4B 6Y4, Canada. matthias.geissler@cnrc-nrc.gc.ca.

|June 16, 2026

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Purification and functional characterization of gag-spike virus-like particles: Process optimization for efficient vaccine production.

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This study presents a novel microfluidic assay for rapid adeno-associated virus (AAV) purification. The automated system efficiently isolates AAV particles from crude lysate in under an hour, improving gene therapy manufacturing.

Area of Science:

Biotechnology
Bioengineering
Molecular Biology

Background:

Adeno-associated virus (AAV) is a critical vector for gene therapy.
Current AAV purification methods can be time-consuming and labor-intensive.
Scalable and efficient purification is essential for AAV manufacturing.

Purpose of the Study:

To develop an integrated, automated microfluidic assay for AAV particle purification.
To optimize the process for speed, efficiency, and compatibility with low-volume samples.
To validate the purification performance, including DNA removal and biological activity.

Main Methods:

Development of a microfluidic cartridge with POROS™ CaptureSelect™ AAVX Affinity Resin.
Utilized a centrifugal platform for pneumatic liquid actuation.

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Integrated 22 automated steps for purification from crude lysate to conditioned AAV sample.

Numerical simulations for flow uniformity and experimental validation with AAV8.

Main Results:

Achieved AAV purification in approximately 30-60 minutes.
Demonstrated higher extraction efficiency compared to manual spin column methods.
Validated plasmid DNA removal exceeding 99.99% via qPCR.
Confirmed biological activity of purified AAV particles in transduction assays.

Conclusions:

The integrated microfluidic assay offers a rapid and efficient method for AAV purification.
The platform addresses limitations in current gene therapy manufacturing, particularly for low-volume production.
This technology has the potential to streamline quality control and accelerate AAV production.