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Abstract

During development of adenovirus-based vaccines, samples have to be analyzed in order to either monitor the production process or control the quality and safety of the product. An important quality attribute is the total concentration of intact adenoviruses, which currently is determined by quantitative polymerase chain reaction (qPCR) or anion exchange-HPLC. Capillary Electrophoresis (CE) was evaluated as alternative to the current methods with the aim to have one single method that allows reliable and fast quantification of adenovirus particles throughout the full process. Intact adenoviruses samples from downstream processing and upstream processing were analyzed directly by CE with UV-detection at 214 nm. Only the samples with high amounts of DNA required a simple sample pretreatment by benzonase. Adenovirus particles were separated from matrix components such as cell debris, residual cell DNA, and/or proteins on a PVA-coated capillary using a BGE consisting of 125 mM Tris, 338 mM tricine and 0.2% v/v polysorbate-20 at pH 7.7. Full factorial design of experiments was used for method optimization as part of the analytical quality by design (AQbD) method development approach. The method was validated for the quantification of adenoviruses on five representative samples from the manufacturing process in the range of 0.5×1011–1.5×1011 adenovirus particles per ml (~80 to 250 pmol/l). The CE method showed intermediate precision of 7.8% RSD on concentration and an accuracy (spiked recovery) of 95–110%. CE proved highly useful for process development support and is being implemented for in-process control testing for adenovirus vaccine manufacturing.

Item Type:	Article
Authors/Creators:	van Tricht, E and Geurink, L and Backus, H and Germano, M and Somsen, GW and Sanger van de Griend, CE
Keywords:	analytical quality by design, capillary electrophoresis, intact adenoviruses, quantification, upstream and downstream processing
Journal or Publication Title:	Talanta
Publisher:	Elsevier Science Bv
ISSN:	0039-9140
DOI / ID Number:	https://doi.org/10.1016/j.talanta.2017.01.013
Copyright Information:	© 2017 Elsevier B.V.
Related URLs:	UTAS Profile: Sanger van de Griend, CE
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