Figure 3.

Optimized AAVX affinity purification process

(A) Process steps of the protocol. (B) Stepwise recovery at each step of the purification process. Vector genomes were quantified via qPCR from aliquots of the sample at each process step and represented as normalized to the lysate. N = 6 biological replicates for both AAV9 and PHP.eB. (C) Overall purification efficiencies of the non-optimized and optimized protocols for AAV9 and PHP.eB combined. Difference was assessed using a two-tailed t test, with ∗p < 0.05. (D) Recovery after filtration + buffer exchange steps for AAV9 and PHP.eB. Note that the values above 100% fall within the range of the approximately 20% precision limit of qPCR titration, and likely do not represent actual recoveries above 100%. (E) Total yields per HYPERFlask across all vectors produced with scAAV9 and scPHP.eB and purified using this protocol. Error bars denote standard deviation in all panels. Note that this includes some vectors that have lower than average production yields. Detailed steps of the purification process are listed in supplemental protocol.