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Related Concept Videos

Downstream Processing01:29

Downstream Processing

Downstream processing begins once fermentation is complete and involves a series of steps to recover and purify products such as acids, vitamins, antibiotics, or proteins.Cell HarvestingFor example, for intracellular protein-based products, the first step is harvesting the cells. This is typically achieved using centrifugation or filtration to separate the cells from the liquid phase.Cell Disruption for Intracellular ProductsIf the target product is intracellular, the harvested cells must be...

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Updated: Jun 26, 2026

Process Development for the Production and Purification of Adeno-Associated Virus AAV2 Vector using Baculovirus-Insect Cell Culture System
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Downstream Process Intensification for AAV Purification by Affinity Chromatography Using Single Pass Tangential Flow

Akshay S Chaubal1, Ronny Horax2,3, Christopher J Yehl4

  • 1Department of Chemical Engineering, Pennsylvania State University, University Park, Pennsylvania.

Biotechnology and Bioengineering
|October 25, 2025
PubMed
Summary
This summary is machine-generated.

Next-generation manufacturing processes for adeno-associated viral vectors (AAV) were intensified using single pass tangential flow filtration (SPTFF). This approach significantly boosted productivity and reduced costs for gene therapy production.

Keywords:
Adeno‐associated virus (AAV)Single pass tangential flow filtrationaffinity chromatographyclarificationcontinuous processingprocess intensification

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Area of Science:

  • Biotechnology
  • Bioprocessing
  • Gene Therapy Manufacturing

Background:

  • Adeno-associated viral vectors (AAV) are crucial for gene therapies, but manufacturing costs limit accessibility.
  • Current AAV downstream processing requires optimization for efficiency and scalability.

Purpose of the Study:

  • To design and evaluate intensified AAV downstream manufacturing processes at bench and pilot scales.
  • To assess the impact of novel clarification and single pass tangential flow filtration (SPTFF) on AAV yield and purity.

Main Methods:

  • Utilized BioOptimal™ MF-SL microfilters and membrane adsorbers for clarification and DNA removal.
  • Implemented SPTFF with regenerated cellulose membranes for AAV concentration.
  • Developed an integrated process combining clarification, SPTFF, and affinity chromatography at pilot scale.

Main Results:

  • Achieved 12X inline volumetric concentration with >99% AAV yield using SPTFF.
  • Demonstrated 48% and 37% host cell protein removal at bench and pilot scales, respectively.
  • The integrated process reduced operating time by 81% and increased productivity 8.5-fold.

Conclusions:

  • SPTFF is a viable technology for intensifying large-scale AAV downstream processing.
  • Intensified processes offer significant improvements in efficiency, cost-effectiveness, and productivity for gene therapy manufacturing.
  • Further development of intensified processes can enhance the affordability and accessibility of AAV-based gene therapies.