Analysis of AAV-Extracted DNA by Charge Detection Mass Spectrometry Reveals Genome TruncationsClick to copy article linkArticle link copied!
- Lauren F. BarnesLauren F. BarnesChemistry Department, Indiana University, 800 E Kirkwood Avenue, Bloomington, Indiana 47405, United StatesMore by Lauren F. Barnes
- Benjamin E. DraperBenjamin E. DraperChemistry Department, Indiana University, 800 E Kirkwood Avenue, Bloomington, Indiana 47405, United StatesMore by Benjamin E. Draper
- Justin KurianJustin KurianAnalytical Research and Development, Pfizer Inc., 875 Chesterfield Pkwy. West, Chesterfield, Missouri 63017, United StatesMore by Justin Kurian
- Yu-Ting ChenYu-Ting ChenAnalytical Research and Development, Pfizer Inc., 875 Chesterfield Pkwy. West, Chesterfield, Missouri 63017, United StatesMore by Yu-Ting Chen
- Tatiana ShapkinaTatiana ShapkinaAnalytical Research and Development, Pfizer Inc., 875 Chesterfield Pkwy. West, Chesterfield, Missouri 63017, United StatesMore by Tatiana Shapkina
- Thomas W. PowersThomas W. PowersAnalytical Research and Development, Pfizer Inc., 875 Chesterfield Pkwy. West, Chesterfield, Missouri 63017, United StatesMore by Thomas W. Powers
- Martin F. Jarrold*Martin F. Jarrold*Email: [email protected]Chemistry Department, Indiana University, 800 E Kirkwood Avenue, Bloomington, Indiana 47405, United StatesMore by Martin F. Jarrold
Abstract

Adeno-associated virus (AAV) is a widely used gene therapy vector. The intact packaged genome is a critical quality attribute and necessary for an effective therapeutic. In this work, charge detection mass spectrometry (CDMS) was used to measure the molecular weight (MW) distribution for the genome of interest (GOI) extracted from recombinant AAV (rAAV) vectors. The measured MWs were compared to sequence masses for a range of rAAV vectors with different GOIs, serotypes, and production methods (Sf9 and HEK293 cell lines). In most cases, the measured MWs were slightly larger than the sequence masses, a result attributed to counterions. However, in a few cases, the measured MWs were significantly smaller than the sequence masses. In these cases, genome truncation is the only reasonable explanation for the discrepancy. These results suggest that direct analysis of the extracted GOI by CDMS provides a rapid and powerful tool to evaluate genome integrity in gene therapy products.
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