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Absolute quantification of dengue virus serotype 4 chimera vaccine candidate in Vero cell culture by targeted mass spectrometry.
Rougemont, Blandine; Simon, Romain; Carrière, Romain; Biarc, Jordane; Fonbonne, Catherine; Salvador, Arnaud; Huillet, Céline; Berard, Yves; Adam, Olivier; Manin, Catherine; Lemoine, Jérôme.
Afiliación
  • Rougemont B; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Simon R; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Carrière R; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Biarc J; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Fonbonne C; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Salvador A; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
  • Huillet C; Sanofi Pasteur, 1541 av. Marcel Mérieux, Marcy l'étoile, France.
  • Berard Y; Sanofi Pasteur, 1541 av. Marcel Mérieux, Marcy l'étoile, France.
  • Adam O; Sanofi Pasteur, 1541 av. Marcel Mérieux, Marcy l'étoile, France.
  • Manin C; Sanofi Pasteur, 1541 av. Marcel Mérieux, Marcy l'étoile, France.
  • Lemoine J; Institut des Sciences Analytiques, UMR 5280 CNRS Université Lyon 1, Université de Lyon, Villeurbanne, France.
Proteomics ; 15(19): 3320-30, 2015 Oct.
Article en En | MEDLINE | ID: mdl-26205729
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Masas / Proteínas Virales / Vacunas Virales / Virus del Dengue Límite: Animals Idioma: En Revista: Proteomics Asunto de la revista: BIOQUIMICA Año: 2015 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Alemania

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría de Masas / Proteínas Virales / Vacunas Virales / Virus del Dengue Límite: Animals Idioma: En Revista: Proteomics Asunto de la revista: BIOQUIMICA Año: 2015 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Alemania