End-to-end characterization of AAV manufacturing process using charge detection mass spectrometry

Charge detection mass spectrometry (CD-MS) has previously been shown to be effective for analyzing predominantly clean bulk drug substances and drug products of recombinant adeno-associated virus (rAAV). CD-MS holds potential for assessing and monitoring the quality attributes of rAAV during the manufacturing process, from harvest to drug product. The mass range of rAAV material has been well characterized experimentally by CD-MS to agree with theoretical values for protein and target genome masses. CD-MS measures the charge range derived from native electrospray ionization of rAAV materials. Ions of rAAV can be separated in data processing using both mass and charge characteristics to improve certainty of rAAV analytics. We demonstrate the capabilities of CD-MS analysis for examining in-process samples from the bulk harvest, affinity chromatography, and anion exchange chromatography. Our results show that the harvest material maintains consistent empty/partial/full ratios when compared to the affinity purified sample. Process impurities were detected for each stage of production and compared to current industry standards. We find that CD-MS is highly reproducible and can be used to effectively track the production process from harvest to bulk drug substance, providing quantitative results at each stage.