Absolute quantification of dengue virus serotype 4 chimera vaccine candidate in Vero cell culture by targeted mass spectrometry

Abstract

Infection by dengue flavivirus is transmitted by mosquitoes and affects tens to hundreds of millions people around the world each year. Four serotypes have been described, all of which cause similar disease. Currently, there no approved vaccines or specific therapeutics for dengue, although several vaccine prototypes are in different stages of clinical development. Among them, a chimeric vaccine, built from the replication machinery of the yellow fever 17D virus, has shown promising results in phase III trials. Accurate quantitation of expressed viral particles in alive attenuated viral antigen vaccine is essential and determination of infectious titer is usually the method of choice. The current paper describes an alternative or orthogonal strategy, namely, a multiplexed and absolute assay of four proteins of the chimera yellow fever/dengue serotype 4 virus using targeted MS in SRM mode. Over 1 month, variability of the assay using a partially purified Vero cell extract was between 8 and 17%, and accuracy was between 80 and 120%. In addition, the assay was linear between 6.25 and 200 nmol/L and could therefore be used in the near future to quantify dengue virus type 4 during production and purification from Vero cells.

Enhancing Dengue Virus Production and Immunogenicity with Celcradle™ Bioreactor: A Comparative Study with Traditional Cell Culture Methods

“…Vero cells, known for their utility in vaccine production due to susceptibility to various viruses [ 18 , 19 , 20 ], are particularly integral to the production of dengue virus vaccines [ 21 , 22 ]. In this study, the adaptation of Vero cells to suspended growth within the CelCradle™ bioreactor system was closely monitored to develop an optimal cell culture protocol that supports continuous viral multiplication.…”

The expanding role of mass spectrometry in the field of vaccine development

“…In a separate study, accurate quantitation of the expressed four viral particles in the tetravalent dengue vaccine (CYD) was performed using targeted MS in selected reaction monitoring (SRM) mode . The study described an orthogonal quantitation strategy (targeted MS in SRM mode) and demonstrated that the variability of the MS method was low (between 8% and 17%) and the assay was linear between 6.25 and 200 nmol/L.…”

Proteome analysis of Duck Tembusu virus (DTMUV)‐infected BHK‐21 cells

“…Emerging technologies, such as proteomic approaches, have become important tools for studying host cellular responses to viral infection and provide specific insights into the cellular mechanisms involved in viral pathogenesis . Several quantitative proteomic approaches have been used to provide specific insights into the gene expression profiles of flaviviruses, including Japanese encephalitis virus (JEV) , Hepatitis C virus (HCV) , and dengue virus (DENV) . These studies have not only increased our understanding of the host response to viral infection but have also shed some light on potential targets for the development of antiviral agents.…”