Human embryonic kidney (HEK) 293 cells and Spodoptera frugiperda (Sf9) insect cells are commonly used to produce adeno-associated virus (AAV)-based gene therapeutics. Read More Below
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Human embryonic kidney (HEK) 293 cells and Spodoptera frugiperda (Sf9) insect cells are commonly used to produce adeno-associated virus (AAV)-based gene therapeutics. However, they are likely to exhibit differences in post translational modifications (PTM) depending on the AAV capsid protein serotype which can impact safety and potency.
Here we explore differences shown in AAV serotype 8 produced in Sf9/baculovirus vs HEK293 cells. Using liquid chromatography mass spectrometry (LC-MS), the relative degrees of modification between proteins from the two expression systems were determined and their purity confirmed. Digestion followed by peptide mapping analysis then identified the exact sites and relative abundances of modified amino acids on the capsid proteins.
Using MS data to improve characterization of AAV from different sources will help scientists to make informed decisions in the development of cost-effective, safe and potent gene therapeutics.
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