Optimized formulation buffer preserves adeno-associated virus-9 infectivity after 4 °C storage and freeze/thawing cycling

Abstract

Adeno-associated virus (AAV) have long been one of the most common and versatile vectors for in vitro and in vivo gene transfer. AAV production protocols are complex and time consuming, one key concern is the recovery and infectivity of viral vector after purification. The buffer used in the storage of AAV at 4 °C and - 80 °C is a crucial factor and methods to improve it have been thoroughly investigated. Viral core facilities have developed formulas using either 0.001% Pluronic F68 or 5% sorbitol in their storage buffers based on the results of this research. Interestingly, few use formulations that include both a non-ionic surfactant and cryopreservative. In this study, AAV9 stored at 4 °C and at - 80 °C in the standard buffers is compared to a buffer that contains 5% glycerol and 0.001% Pluronic F68. By viral genome quantitation with qPCR, all three formulations show the same extent of viral titer loss at 4 °C, while after several cycles of freeze/thaws at - 80 °C, the viral recovery and infectivity in the preparation with both glycerol and Pluronic F68 was most stable compared to the other buffers.

Keywords: AAV; Formulation buffer; Glycerol; Pluronic F68; Sorbitol.