Compositions and methods to prevent AAV vector aggregation

摘要

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. μ˜500 mM) 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 6.4×1013 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.

主权项

1. A composition for the storage of purified, recombinant adeno-associated virus (AAV) vector particles, comprising:
purified, recombinant AAV vector particles at a concentration exceeding 1×1013 vg/ml up to 6.4×1013 vg/ml; a pH buffer, wherein the pH of the composition is between 7.5 and 8.0; and excipients comprising one or more multivalent ions selected from the group consisting of citrate, sulfate, magnesium, and phosphate; wherein the ionic strength of the composition is greater than 200 mM, and wherein the purified AAV vector particles are stored in the composition without significant aggregation.