Related Experiment Video
Updated: Sep 12, 2025
10:31
Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System
Published on: January 13, 2022
9.6K
Harvesting AAV by tangential flow filtration using reverse asymmetric membranes
Xiaolei Hao1, Ronny Horax2, Xianghong Qian1
1Department of Biomedical Engineering, University of Arkansas, Fayetteville, Arkansas, USA.
Biotechnology Progress
|August 4, 2025
Summary
This study demonstrates reverse asymmetric hollow fiber membranes effectively harvest adeno-associated virus (AAV). Optimized flow rates and transmembrane pressure ensure high permeability and 94% virus recovery during bioreactor clarification.
Area of Science:
- Biotechnology
- Membrane Filtration
- Virology
Background:
- Efficient bioreactor clarification for virus particle harvesting is a significant challenge in biopharmaceutical production.
- Tangential flow filtration (TFF) offers a scalable solution adaptable to various bioprocessing operations.
Purpose of the Study:
- To investigate the feasibility of using reverse asymmetric hollow fiber membranes for harvesting adeno-associated virus serotype 2 (AAV2).
- To determine the critical flux and optimize operating parameters for stable and efficient AAV2 recovery.
Main Methods:
- Utilized reverse asymmetric hollow fiber membranes with the open support structure facing the feed stream.
- Conducted flux stepping experiments in total recycle mode to determine critical flux.
- Performed AAV2 harvesting at a wall shear rate of 2000 s⁻¹ and a permeate flux of 15 Lm⁻²h⁻¹.
Main Results:
- Critical flux increased with increasing wall shear rate, indicating improved membrane stability at higher flow rates.
- A stable transmembrane pressure of 3.5 kPa was maintained during concentration and diafiltration.
- Achieved a high virus recovery rate of 94% with minimal product dilution.
Conclusions:
- Reverse asymmetric hollow fiber membranes are effective for AAV2 harvesting, offering a stable and permeable secondary membrane.
- Optimized operating conditions, including wall shear rate and transmembrane pressure, are crucial for maximizing virus recovery and process efficiency.
