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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.

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|May 18, 2026
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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.

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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.