Takara AAVpro Extraction Solution \1 Set

Isolation of AAV particles from AAV particle-producing cells is conventionally performed using the freeze-thaw or sonication methods. However, these methods are time consuming and require special equipment. The AAVpro Extraction Solution is a reagent for the extraction of AAV particles from AAV particle-producing cells. This reagent provides a simple and efficient method for AAV particle isolation; the reagents are added to virus-producing cells and viral particles are recovered by centrifugation. The extracted AAV particle solution contains only a small amount of host protein and nucleic acid contamination. The resulting viral extract is well-suited for cell infection or further purification of viral particles.

Comparison of AAV Extraction Solution with the freeze-thaw method.The titer of AAV2 extract solutions was determined by real-time PCR (vector genome; vg). Then, the equivalent of 1x109vg of the AAV2 extract solution was analyzed by SDS-PAGE to evaluate protein impurity (Panel A). In addition, residual dsDNA content was assayed using the intercalation method (Panel B). The use of the AAV Extraction Solution clearly reduced the amount of protein impurities and dsDNA in comparison with the freeze-thaw method.

Extraction of serotype 1, 2, and 6 AAV particles.AAV particles were extracted from HEK 293 cells using either the AAVpro Extraction Solution or the freeze-thaw method. The AAVpro Titration Kit (for Real Time PCR) Ver. 2 (Cat. # 6233) was used to measure the titer of the AAV solutions. The results indicate that the AAVpro Extraction Solution can be used to efficiently extract AAV particles that are serotype 1, 2, or 6.