High-recovery AAV clarification using a multiplexed spiral inertial microfluidic platform.
Alexander Bevacqua1,2,3, Do Hyun Park2, Sheryar Khan4
1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. jyhan@mit.edu.
Lab on a Chip
|May 18, 2026
Summary
This study introduces spiral inertial microfluidics for AAV clarification, achieving 85% viral vector recovery. Combining this with depth filtration enhances throughput and overall recovery in bioprocessing.
Area of Science:
- Biotechnology
- Bioprocessing
- Microfluidics
Background:
- Cell and gene therapies require efficient viral vector production.
- Current AAV purification uses membrane filters, leading to clogging and product loss.
- Developing advanced clarification methods is crucial for clinical adoption.
Purpose of the Study:
- To evaluate spiral inertial microfluidic technology as a membraneless AAV clarification method.
- To assess the efficiency of AAV recovery and cell removal using this microfluidic system.
- To investigate the integration of microfluidics with depth filtration for improved bioprocessing.
Main Methods:
- A parallelized plastic spiral microfluidic system was developed for cell clarification.
- The system processed cell culture at 20 mL/min, separating AAVs from host cells.
- A two-step process combining microfluidics with 3M Harvest RC depth filters was implemented.
Main Results:
- The microfluidic platform achieved 85% viral vector recovery but incomplete cell removal (up to 20% cells in harvest).
- Integrating microfluidics before depth filtration increased filter throughput by 56.25% (44.8 to 70 L/m²).
- The two-step process yielded approximately 74% AAV recovery, surpassing typical clarification efficiencies.
Conclusions:
- Spiral inertial microfluidics offers a promising alternative for AAV clarification.
- Combining microfluidics with depth filtration significantly enhances bioprocessing efficiency and recovery.
- Further optimization and scale-out of microfluidic systems could boost overall AAV production recovery.
