Development of purification steps for several AAV serotypes using POROS™ CaptureSelect™ AAVX affinity chromatography

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Innovator Insight

M Toueille, L Dejoint, E Attebi, J Cartigny, C Rasle, S Potier, S Rundwasser, L Guianvarc’h, C Lebec & M Hebben

latest advances in BIOPROCESSING: overcoming bottlenecks

Genethon is an integrated R&D centre created in 1990 by the French Association of Muscular Dystrophy. The Centre performs both gene therapy translational research and clinical development, with a specific focus on rare genetic diseases such as neuromuscular disorders, immuno-deficiencies, eye diseases and liver diseases. The two main vector platforms used at Genethon are AAV and lentiviral vectors.

Published: Sept 18 2018
DOI: 10.18609/cgti.2018.061
Citation: Cell Gene Therapy Insights 4(7), 637-645
Open access

Current purification methods for viral vector manufacturing usually involve multiple steps of chromatography, for example ion exchange and hydrophobic interaction, which leads to cumulative yield loss and ultimately, increased cost.

Thermo Fisher’s CaptureSelect™ technology addresses this issue by providing high-affinity binding ligands for affinity chromatography. This approach provides high purity and productivity in fewer steps than traditional purification processes, while also offering process consistency and scalability.

The technology is based on the utilisation of the variable domains of camelid heavy-chain-only antibodies (VHH) which offer full functionality in antigen-specific recognition and also high-affinity binding (the three CDR regions of the VHH ligand provide unique, tunable specificity). Due to their compact structure, the domains are highly robust and capable of withstanding the various conditions typical of chromatography runs. Additionally, the selected ligands have the advantage of being produced in an animal origin-free system - Saccharomyces cerevisiae (baker’s yeast).

The result is a unique screening technology for target specificity, mild elution and stability, which is designed for commercial purification processes.

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