Intensifying Animal Cell Culture for Virus Vaccine Manufacturing

强化动物细胞培养以生产病毒疫苗

• 批准号: 519884-2017
• 负责人: AUCOIN, Marc
• 金额: $ 11.47万
• 依托单位: University of Waterloo
• 依托单位国家: 加拿大
• 项目类别: Collaborative Research and Development Grants
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=644296
• 关键词: Intensifying; Animal; Cell; Culture; Virus

Past challenges of vaccine manufacturing have arguably been related to scale-up, and for the most part the challenge has been met. Today, the unmet challenges, which have dogged the vaccine industry for the past 30 years, are the lack of manufacturing flexibility, especially in cases of sudden virus outbreaks, and the failure to intensify the production of viruses. Three major intensification strategies include: the use of cell lines for virus propagation instead of embryonated eggs or mouse brains; the adaptation of cell lines to suspension that can enable high density cultures; and the move from batch to continuous modes of operating the process. In this project, Profs Aucoin and Kamen, who have a long history of working collaboratively together, are partnering with Sanofi, a leading international vaccine manufacturer, to transform the Vero cell culture platform into a leading edge technology amenable to intensification. There is a clear need and demand for efficient, large scale production of vaccines to prevent disease related to viral infection in the world, evidenced by the fact that today's vaccine market is worth ~25 billion dollars and is expected to more than double by 2020. In the last two decades, four viruses have been the documented cause of global epidemics: SARS coronavirus; avian and swine influenza viruses; Ebola virus; and recently, Zika virus. Cell culture is the most attractive manufacturing platform for rapid, scalable and controlled production of viral vaccines - and Vero is the most widely accepted platform for viral vaccine manufacturing. Despite reaching significant intensification in Chinese Hamster Ovary cultures for the production of monoclonal antibodies, productive infections at high cell densities in cell culture for recombinant protein production or virus manufacture remains elusive. The goals of this work are to enhance the genetic understanding of the Vero cell line; determine the genetic and metabolic underpinnings of the peak cell density effect and the phenotypic transformation from adherent to suspension cell state; and develop a (semi)-continuous mode of operation that enables higher volumetric productivity.******

过去疫苗生产的挑战可以说与扩大规模有关，而且大部分挑战已经得到解决。如今，过去30年来一直困扰疫苗行业的未解决的挑战是缺乏生产灵活性，特别是在病毒突然爆发的情况下，以及未能加强病毒的生产。三种主要的强化策略包括：使用细胞系代替胚胎卵或小鼠大脑进行病毒繁殖；使细胞系适应悬浮，从而实现高密度培养；以及从间歇操作模式转变为连续操作过程模式。在这个项目中，有着长期合作历史的 Aucoin 和 Kamen 教授与国际领先的疫苗制造商赛诺菲合作，将 Vero 细胞培养平台转变为适合集约化的前沿技术。世界上显然需要高效、大规模生产疫苗来预防与病毒感染相关的疾病，当今的疫苗市场价值约 250 亿美元，预计到 2020 年将增加一倍多，这一事实证明了这一点。在过去二十年中，有四种病毒已被记录为全球流行病的原因：SARS 冠状病毒；禽流感和猪流感病毒；埃博拉病毒；最近，寨卡病毒。细胞培养是快速、可扩展和受控生产病毒疫苗的最具吸引力的制造平台，而 Vero 是最广泛接受的病毒疫苗制造平台。尽管在中国仓鼠卵巢培养物中实现了单克隆抗体生产的显着强化，但在细胞培养物中以高细胞密度进行重组蛋白生产或病毒制造的有效感染仍然难以实现。这项工作的目标是增强对 Vero 细胞系的遗传理解；确定峰值细胞密度效应和从贴壁细胞状态到悬浮细胞状态的表型转变的遗传和代谢基础；并开发一种（半）连续操作模式，以实现更高的容积生产率。******