Analysis of residual DNA by high-throughput sequencing (SSV-Seq) in rAAV lots

With the success of clinical trials using recombinant adeno-associated viral vectors (rAAV), regulatory agencies ask for a more comprehensive characterization of process- and product- related impurities found in rAAV stocks in order to assess the potential risks for patients. The CPV (translational vector core) has developed a protocol, the Single-Stranded DNA Virus Sequencing (SSV-Seq) ^1-3 and a dedicated bioinformatics pipeline in order to :

Identify and Quantify residual DNAs (relative percentage of each DNA species)
Analyze rAAV genome identity (including SNV)

The CPV offers to academics and biotech companies to perform this non-selective quality control for a comprehensive analysis of residual DNAs in your purified rAAV batches by high-throughput sequencing.

The CPV quality management system meets the requirements of the Management System Standards ISO 9001:2015.

1. Lecomte, E, Tournaire, B, Cogné, B, Dupont, J-B, Lindenbaum, P, Martin-Fontaine, M, et al. (2015). Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded Virus Next-generation Sequencing. Mol. Ther. Nucleic Acids 4: e260.

2. Penaud-Budloo, M, Lecomte, E, Guy-Duché, A, Saleun, S, Roulet, A, Lopez-Roques, C, et al. (2017). Accurate Identification and Quantification of DNA Species by Next-Generation Sequencing in Adeno-Associated Viral Vectors Produced in Insect Cells. Hum. Gene Ther. Methods 28: 148–16

3. Lecomte, E, Leger, A, Penaud-Budloo, M, Ayuso, E (2019). Single-Stranded DNAVirus Sequencing (SSV-Seq) for Characterization of Residual DNA and AAV Vector Genomes, Methods Mol Biol. 2019;1950:85-106.

Mis à jour le 16 octobre 2020.