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Measurement of Adenovirus-Based Vector Heterogeneity

Hickey, John M; Jacob, Shaleem I; Tait, Andrew S; Vahid, Fatemeh Dastjerdi; Barritt, Joseph; Rouse, Sarah; Douglas, Alexander; ... Bracewell, Daniel G; + view all (2022) Measurement of Adenovirus-Based Vector Heterogeneity. Journal of Pharmaceutical Sciences 10.1016/j.xphs.2022.12.012. (In press). Green open access

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Abstract

Adenovirus vectors have become an important class of vaccines with the recent approval of Ebola and COVID-19 products. In-process quality attribute data collected during Adenovirus vector manufacturing has focused on particle concentration and infectivity ratios (based on viral genome: cell-based infectivity), and data suggest only a fraction of viral particles present in the final vaccine product are efficacious. To better understand this product heterogeneity, lab-scale preparations of two Adenovirus viral vectors, (Chimpanzee adenovirus (ChAdOx1) and Human adenovirus Type 5 (Ad5), were studied using transmission electron microscopy (TEM). Different adenovirus morphologies were characterized, and the proportion of empty and full viral particles were quantified. These proportions showed a qualitative correlation with the sample's infectivity values. Liquid chromatography-mass spectrometry (LC-MS) peptide mapping was used to identify key adenovirus proteins involved in viral maturation. Using peptide abundance analysis, a ∼5-fold change in L1 52/55k abundance was observed between low-(empty) and high-density (full) fractions taken from CsCl ultracentrifugation preparations of ChAdOx1 virus. The L1 52/55k viral protein is associated with DNA packaging and is cleaved during viral maturation, so it may be a marker for infective particles. TEM and LC-MS peptide mapping are promising higher-resolution analytical characterization tools to help differentiate between relative proportions of empty, non-infectious, and infectious viral particles as part of Adenovirus vector in-process monitoring, and these results are an encouraging initial step to better differentiate between the different product-related impurities.

Type: Article
Title: Measurement of Adenovirus-Based Vector Heterogeneity
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.xphs.2022.12.012
Publisher version: https://doi.org/10.1016/j.xphs.2022.12.012
Language: English
Additional information: © 2022 Published by Elsevier Inc. on behalf of American Pharmacists Association. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Keywords: Adenovirus-based Vaccine, Analytical Characterization, Critical Quality Attributes, In-process testing, Mass Spectrometry, Transmission Electron Microscopy
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10162601
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