Production and purification of adeno-associated virus (AAV) at the 50 L scale

To build upon previous work demonstrating the scalability of the Gibco™ CTS™ AAV-MAX Helper-Free AAV Production System to produce AAV6-GFP in the Thermo Scientific™ DynaDrive™ Single-Use Bioreactor (S.U.B.)[1], here we provide data on the production, purification, and characterization of an AAV6 anti-CD19 chimeric antigen receptor (AAV6-CD19CAR) in the DynaDrive S.U.B. at the 50 L scale. Two DynaDrive S.U.B.s were run in parallel with shake flask controls to demonstrate scalability of the process using a non-GFP gene of interest. Furthermore, we provide guidance for a scalable downstream process for purification of the 50 L AAV6-CD19CAR. This purification strategy utilizes depth filters for supernatant clarification and tangential flow filtration (TFF) for concentration and buffer exchange prior to loading of the AAV onto the Thermo Scientific™ POROS™ CaptureSelect™ AAVX Affinity Resin. Utilizing this process, we obtained and purified a biologically active AAV6-CD19CAR, demonstrating a feasible and further scalable end-to-end workflow.
In all steps of the process, Gibco™ CTS™ Viral Production Cells 2.0 (VPC 2.0) were maintained in Gibco™ CTS™ Viral Production Medium containing 4 mM Gibco™ GlutaMAX™ Supplement (Table 1). The cells were thawed and maintained in shake flasks as recommended in the AAV-MAX system user guide [2] before inoculating each of the bioreactors at a low turndown ratio to further expand the cells prior to each production run.

Fill out this form to download

Copyright © 2023 Content Lead, All Rights Reserved.