Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers

Dengue is a mosquito-borne disease and is a major global health threat, especially in tropical and subtropical regions. In this study, we describe the optimization of virus production for a dengue live-attenuated quadrivalent vaccine candidate and demonstrate scalability of a stirred tank bioreactor...

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Main Authors: Shannon Haughney, Seth Clark, Elizabeth Carey, Nelson Lee Afanador, Victoria Stabile, Bernard Kang, Samantha Marrone, Jillian Shingler, Carl Hofmann, Christopher J. Wang, Christopher Ton
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Vaccine: X
Online Access:http://www.sciencedirect.com/science/article/pii/S259013622500052X
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author Shannon Haughney
Seth Clark
Elizabeth Carey
Nelson Lee Afanador
Victoria Stabile
Bernard Kang
Samantha Marrone
Jillian Shingler
Carl Hofmann
Christopher J. Wang
Christopher Ton
author_facet Shannon Haughney
Seth Clark
Elizabeth Carey
Nelson Lee Afanador
Victoria Stabile
Bernard Kang
Samantha Marrone
Jillian Shingler
Carl Hofmann
Christopher J. Wang
Christopher Ton
author_sort Shannon Haughney
collection DOAJ
description Dengue is a mosquito-borne disease and is a major global health threat, especially in tropical and subtropical regions. In this study, we describe the optimization of virus production for a dengue live-attenuated quadrivalent vaccine candidate and demonstrate scalability of a stirred tank bioreactor serum-free microcarrier process. We utilized a design-of-experiment (DoE) methodology to optimize for pH and temperature and evaluate effect of Multiplicity of Infection (MOI) and Time of Infection (TOI) for each serotype during virus production in 2 L and 3 L bioreactors. The optimal pH and temperature for dengue virus production achieved ∼10× higher virus titer at peak virus production for all 4 serotypes compared to baseline condition. Different MOI and TOI did not impact peak virus production. Results from the DoE studies were successfully scaled to the 50 L Single-Use Bioreactor (SUB).
format Article
id doaj-art-564ad35319a244e0a3a20d7dc2d7c8a2
institution OA Journals
issn 2590-1362
language English
publishDate 2025-06-01
publisher Elsevier
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series Vaccine: X
spelling doaj-art-564ad35319a244e0a3a20d7dc2d7c8a22025-08-20T02:30:58ZengElsevierVaccine: X2590-13622025-06-012410065810.1016/j.jvacx.2025.100658Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriersShannon Haughney0Seth Clark1Elizabeth Carey2Nelson Lee Afanador3Victoria Stabile4Bernard Kang5Samantha Marrone6Jillian Shingler7Carl Hofmann8Christopher J. Wang9Christopher Ton10Bioprocess Drug Substance Commercialization, Merck & Co., Inc., Rahway, NJ, USABiostatistics and Research Decision Sciences, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USABiostatistics and Research Decision Sciences, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USAAnalytical Research & Development, Merck & Co., Inc., Rahway, NJ, USAAnalytical Research & Development, Merck & Co., Inc., Rahway, NJ, USAProcess Research & Development, Merck & Co., Inc., Rahway, NJ, USA; Corresponding author.Dengue is a mosquito-borne disease and is a major global health threat, especially in tropical and subtropical regions. In this study, we describe the optimization of virus production for a dengue live-attenuated quadrivalent vaccine candidate and demonstrate scalability of a stirred tank bioreactor serum-free microcarrier process. We utilized a design-of-experiment (DoE) methodology to optimize for pH and temperature and evaluate effect of Multiplicity of Infection (MOI) and Time of Infection (TOI) for each serotype during virus production in 2 L and 3 L bioreactors. The optimal pH and temperature for dengue virus production achieved ∼10× higher virus titer at peak virus production for all 4 serotypes compared to baseline condition. Different MOI and TOI did not impact peak virus production. Results from the DoE studies were successfully scaled to the 50 L Single-Use Bioreactor (SUB).http://www.sciencedirect.com/science/article/pii/S259013622500052X
spellingShingle Shannon Haughney
Seth Clark
Elizabeth Carey
Nelson Lee Afanador
Victoria Stabile
Bernard Kang
Samantha Marrone
Jillian Shingler
Carl Hofmann
Christopher J. Wang
Christopher Ton
Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
Vaccine: X
title Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
title_full Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
title_fullStr Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
title_full_unstemmed Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
title_short Optimization for the production of a dengue live-attenuated Quadrivalent vaccine in Vero cells grown on microcarriers
title_sort optimization for the production of a dengue live attenuated quadrivalent vaccine in vero cells grown on microcarriers
url http://www.sciencedirect.com/science/article/pii/S259013622500052X
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