title: Simple downstream process based on detergent treatment improves yield and in vivo transduction efficacy of adeno-associated virus vectors.
abstract: Recombinant adeno-associated viruses (rAAV) are promising candidates for gene therapy approaches. The last two decades were particularly fruitful in terms of processes applied in the production and purification of this type of gene transfer vectors. This rapid technological evolution led to better yields and higher levels of vector purity. Recently, some reports showed that rAAV produced by transient tri-transfection method in adherent human embryonic kidney 293 cells can be harvested directly from supernatant, leading to easier and faster purification compared to classical virus extraction from cell pellets. Here, we compare these approaches with new vector recovery method using small quantity of detergent at the initial clarification step to treat the whole transfected cell culture. Coupled with tangential flow filtration and iodixanol-based isopycnic density gradient, this new method significantly increases rAAV yields and conserves high vector purity. Moreover, this approach leads to the reduction of the total process duration. Finally, the vectors maintain their functionality, showing unexpected higher in vitro and in vivo transduction efficacies. This new development in rAAV downstream process once more demonstrates the great capacity of these vectors to easily accommodate to large panel of methods, able to furthermore ameliorate their safety, functionality, and scalability.
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Dias Florencio
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Precigout
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Cyriaque
authors.initials: C
authors.lastname: Beley
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Buclez
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Luis
authors.initials: L
authors.lastname: Garcia
authors.affiliation: U1179 UVSQ-INSERM, Université de Versailles Saint-Quentin en Yvelines, UFR des Sciences de la Santé, Equipe Biothérapies des Maladies Neuromusculaires , Montigny-le-Bretonneux, France.
authors.forename: Rachid
authors.initials: R
authors.lastname: Benchaouir