Microcarrier culture system for human influenza vaccine

人流感疫苗微载体培养系统

• 批准号: 6689839
• 负责人: William J Hillegas
• 金额: $ 38.79万
• 依托单位: SOLOHILL ENGINEERING, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689839
• 关键词: MDCK cell; Vero cells; bioreactors; biotechnology; cost effectiveness; drug design /synthesis /production; influenza vaccines; influenzavirus A; polystyrenes; technology /technique development; tissue /cell culture; trypsin; vaccine development

DESCRIPTION (provided by applicant): The goal of the proposed research is to develop an optimized microcarrier-bioreactor cell culture system that enables the rapid, efficient, large scale, low cost production of human influenza vaccine. Most human influenza virus vaccine is produced in embryonated eggs. The Federal government and other worldwide organizations are seeking alternative cell-culture approaches to influenza vaccine production since there are inherent difficulties with the egg-derived vaccines. Under normal conditions there are long lead times between (a) the identification of particular virus strains likely in each flu season and (b) the availability of appropriate vaccines against these virus strains. Microcarrier-bioreactor cell-culture technology has the potential to lower production costs and reduce the time required to produce 93 million doses of monovalent vaccine to a few months. This potential to rapidly achieve large scale, would be invaluable in case of an influenza pandemic and possibly could counter a pandemic or bioterrorist acts. To achieve the research goals, five specific aims will optimize microcarrier-bioreactor culture process parameters that affect viral yield with Vero and MDCK cells and develop protocols applicable to economic commercial use. We will (i) license manufacturing protocols to large producers of flu vaccine and (ii) sell Hillex microcarriers for those protocols. Specific Aim I. To identify and integrate critical cell culture and viral growth parameters for optimal production of human influenza virus in Vero cells grown in Hillex microcarrier-suspension culture. Specific Aim II. To extend the range of viral growth parameters (as identified in Specific Aim I) to include six additional strains of influenza virus and to compare the virus production in direct "head-to-head" competition between embryonated eggs and Vero cells grown in Hillex microcarrier-suspension culture. Specific Aim III. To assess production of human influenza virus in Vero cells grown in a microcarrier-bioreactor culture at the one liter scale. Specific Aim IV: To develop a process to expand and scale-up Vero cells from a working seed ampoule to a five liter culture and to demonstrate that the passaged cells will support high influenza virus concentrations at a larger scale. Specific Aim V: To identify and integrate critical cell culture parameters for optimal production of human influenza virus (A/Wuhan/395/95) in MDCK cells grown in Hillex microcarrier-bioreactor culture.

描述（由申请方提供）：拟定研究的目标是开发一种优化的微载体-生物反应器细胞培养系统，能够快速、高效、大规模、低成本生产人流感疫苗。大多数人流感病毒疫苗是在含胚鸡蛋中生产的。联邦政府和其他世界性组织正在寻求流感疫苗生产的替代细胞培养方法，因为蛋源疫苗存在固有的困难。在正常情况下，在（a）鉴定每个流感季节可能出现的特定病毒株与（B）获得针对这些病毒株的适当疫苗之间有很长的准备时间。微载体生物反应器细胞培养技术有可能降低生产成本，并将生产9300万剂单价疫苗所需的时间缩短到几个月。这种迅速实现大规模的潜力在流感大流行的情况下将是非常宝贵的，并且可能可以对抗大流行或生物恐怖主义行为。 为了实现研究目标，五个具体目标将优化影响Vero和MDCK细胞病毒产量的微载体生物反应器培养工艺参数，并开发适用于经济商业用途的方案。我们将（i）向流感疫苗的大型生产商许可生产协议，（ii）为这些协议销售Hillex微载体。 具体目标一。确定并整合关键细胞培养和病毒生长参数，以在Hillex微载体悬浮培养中生长的Vero细胞中最佳生产人流感病毒。 具体目标二。扩大病毒生长参数的范围（如特定目标I中所确定），以包括另外6种流感病毒株，并比较在Hillex微载体悬浮培养物中生长的鸡胚和Vero细胞之间直接“头对头”竞争的病毒产量。 具体目标三。评估在1 L规模微载体生物反应器培养物中生长的Vero细胞中人流感病毒的产量。 具体目标四：开发一种将Vero细胞从工作种子细胞扩增和扩大到5升培养物的工艺，并证明传代细胞将在更大规模下支持高流感病毒浓度。 具体目标五：确定并整合在Hillex微载体-生物反应器培养物中生长的MDCK细胞中人流感病毒（A/Wuhan/395/95）最佳生产的关键细胞培养参数。