Summary
Along with the wide use of lentiviral vectors (LVVs) in cell and gene therapies, there comes the increasing requirement for accurate and robust analytical methods to validate the potency and quality of the vectors produced.
We implemented several alterations to our ITA assay with the goal to significantly improve throughout, without compromising data accuracy and precision:
We improved the assay by successfully scaling down the plate format, and by onboarding a more convenient DNA extraction method.
We attempted to implement primers which bind specifically to integrated sequence. However, the performance of these primers was not as expected so we were unable to eliminate the passage step as planned. Further work is required to determine the cause of the background amplification observed post-transduction.
Overall, these changes have successfully improved the throughput and data quality of our ITA assay, which will positively impact the analytics process downstream from our LV productions.