Compositions and methods to prevent aav vector aggregation

Details Compositions and methods to prevent aav vector aggregation Compositions and methods to prevent aav vector aggregation

C - Chemistry, Metallurgy

12

N

C12N 7/02 (2006.01)

Patent

CA 2569244

Compositions and methods are provided for preparation of concentrated stock solutions of AAV virions without aggregation. Formulations for AAV preparation and storage are high ionic strength solutions (e.g. ~ ~500mM) that are nonetheless isotonic with the intended target tissue. This combination of high ionic strength and modest osmolarity is achieved using salts of high valency, such as sodium citrate. AAV stock solutions up to 6.4x1013 vg/mL are possible using the formulations of the invention, with no aggregation being observed even after ten freeze-thaw cycles. The surfactant Pluronic® F68 may be added at 0.001% to prevent losses of virions to surfaces during handling. Virion preparations can also be treated with nucleases to eliminate small nucleic acid strands on virions surfaces that exacerbate aggregation.

L'invention concerne des compositions et des méthodes de préparation de solutions mères concentrées de virions AAV sans agrégation. Les formulations de préparation et de stockage d'AAV sont des solutions à force ionique élevée (par exemple ? ~500mM) qui sont néanmoins isotoniques avec le tissu ciblé. Cette combinaison de force ionique élevée et d'osmolarité réduite est obtenue à l'aide de sels à valence élevée, tels que le citrate de sodium. Les formulations de l'invention permettent d'obtenir des solutions mères d'AAV allant jusqu'à 6,4x1013 vg/mL sans agrégation observée y compris après dix cycles de congélation-décongélation. L'agent de surface Pluronic® F68 peut être ajouté à 0,001 % afin de prévenir les pertes de virions en surface au cours de la manipulation. Les préparations de virions peuvent également être traitées à l'aide de nucléases pour éliminer les petits brins d'acides nucléiques sur les surfaces des virions exacerbant l'agrégation.

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