2021
DOI: 10.1101/2021.09.02.458670
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Assessment of genome packaging in AAVs using Orbitrap-based charge detection mass spectrometry

Abstract: Adeno-associated viruses (AAV) represent important gene therapy vectors with several approved clinical applications and numerous more in clinical trials. Genome packaging is an essential step in the bioprocessing of AAVs and needs to be tightly monitored to ensure the proper delivery of transgenes and the production of effective drugs. Current methods to monitor genome packaging have limited sensitivity, a high demand on labour, and struggle to distinguish between packaging of the intended genome or unwanted s… Show more

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Cited by 3 publications
(6 citation statements)
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“…Overall, the CD-MS results support the integrity of transgene packaging in these rAAV samples, with no evidence of truncated gene products (which would manifest as intermediate masses between the empty and filled populations) in these preparations. 37 These results highlight the utility of CD-MS measurements to probe transgene-related rAAV attributes.…”
Section: Resultsmentioning
confidence: 82%
See 2 more Smart Citations
“…Overall, the CD-MS results support the integrity of transgene packaging in these rAAV samples, with no evidence of truncated gene products (which would manifest as intermediate masses between the empty and filled populations) in these preparations. 37 These results highlight the utility of CD-MS measurements to probe transgene-related rAAV attributes.…”
Section: Resultsmentioning
confidence: 82%
“…43 The relative ion abundances of empty and filled capsids detected by CD-MS have previously shown excellent quantitative accuracy across preparations. 37…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…[8][9] However, CD-MS analysis of AAVs often requires relatively high concentrations of sample in the range of 5-100×10 12 capsids per ml. [10][11] More broadly, native MS is also limited in sensitivity and usually requires approximately micromolar concentrations of analyte. 12 A comparable method, mass photometry, has lower nanomolar levels of detection for protein complexes.…”
Section: Introductionmentioning
confidence: 99%
“…In addition, the SedFit data processing software is not 21 CFR Part 11 compliant, further complicating the validation of AUC for routine GMP testing (8). Charge detection mass spectrometry (CDMS) is still in its infancy and carries the weight of validation challenges in GMP and QC environments (9,10). Transmission electron microscopy with negative staining (TEM) is also commonly used to identify and count empty and full capsids.…”
Section: Introductionmentioning
confidence: 99%