Microcarrier / CEF / Media SYSTEM to make Viral Vaccines

用于制造病毒疫苗的微载体/CEF/培养基系统

• 批准号: 6534229
• 负责人: William J Hillegas
• 金额: $ 39.61万
• 依托单位: SOLOHILL ENGINEERING, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6534229
• 关键词: Herpesviridae vaccine; bioreactors; biotechnology; chick embryo; drug design /synthesis /production; fibroblasts; mass tissue /cell culture; technology /technique development; vaccine development

DESCRIPTION (Provided by Applicant): This research plan is to develop a Microcarrier-Bioreactor Cell Culture System for the growth and efficient recovery of infected, whole cells from microcarriers in controlled, bioreactor cultures. Primary chick embryo fibroblast (CEF) cells are widely used in viral vaccine production (for both avian and human vaccines), but there are no known licensed-manufacturing protocols which utilize a Microcarrier-Bioreactor System for this purpose. Focus will be on the production of a whole cell Marek's Disease vaccine for avian applications. The System technology developed in the course of these studies could replace large-scale industrial processes in which (1) primary CEF cells are utilized in roller bottles, (2) embryonated, fertile eggs function as tiny bioreactors, such as in the manufacturing of human influenza vaccine or (3) whole cell vaccines and/or immunotherapies. Specific Aims I, II and VI. To optimize cell culture conditions for a one to two day HVT-production run using primary CEF cells, secondary CEF cells and immortalized CEF-like cells at high density, in a one-liter bioreactor culture. Specific Aim III. To optimize culture conditions for a three-day HVT-production run using secondary CEF cells at the one-liter scale in bioreactor culture. Specific Aim IV & V. To develop protocols for rapid harvesting of HVT-infected cells, and separation of those cells from microcarriers, and to achieve high-titer Marek's vaccine production. PROPOSED COMMERCIAL APPLICATION: Many animal vital vaccines (e.g. Marek's) and human viral vaccines are produced using chick embryo cells, either in roller bottle systems or in embryonated chicken eggs. Neither process uses microcarrier - bioreactor systems. Large scale production of Marek's vaccine in a Miaocarrier-Bioreactor System, will lead to substantial economic advantages when bioreactors are compared to old technology. Product quality and consistency should also be improved. and manufacturing costs reduced. The System herein proposed will utilize (1) primary or secondary CEF cells, or immortalized cell lines. (2) special media formulations, (3) bovine sera and will evolve the (4) protocols to produce vaccines in this System. The technology should extrapolate to other whole-cell human vaccines. PROPOSED COMMERCIAL APPLICATION: The current short supply of human influenza vaccine serves to underline the importance for developing technology that is an alternative to embryonated egg production methods. The lead time from (1) a CDC estimate of the specific influenza virus variant likely to strike the USA, to (2) the availability of the vaccine on a large scale is exceedingly long, many months. In the event of an influenza pandemic this many-month lead-time could result in a high fatality rate reminiscent of the 1917-18 pandemic. If influenza virus vaccine can be produced in very large-scale bioreactors, with a fast turn around time (a couple months), the high fatality rate of a pandemic might be averted. The research proposed has the potential to address this issue.

描述（由申请人提供）：本研究计划旨在开发一种 微载体-生物反应器细胞培养系统 在受控的生物反应器中从微载体回收感染的全细胞 cultures.原代鸡胚成纤维细胞（CEF）广泛用于病毒感染。 疫苗生产（禽流感和人类疫苗），但没有已知的 使用微载体-生物反应器系统的许可生产方案 用于此目的。重点将放在生产一个完整的细胞马雷克的 禽流感疫苗。系统技术的发展， 这些研究过程可以取代大规模的工业过程， (1)在滚瓶中使用原代CEF细胞，（2）含胚的、可育的 鸡蛋的功能是微型生物反应器，例如在制造人类 流感疫苗或（3）全细胞疫苗和/或免疫疗法。 具体目标一、二和六。为了优化细胞培养条件， 使用原代CEF细胞、二代CEF细胞和 在1升生物反应器培养物中以高密度培养永生化CEF样细胞。 具体目标三。为了优化三天HVT生产的培养条件， 在生物反应器培养中以1升规模使用次级CEF细胞运行。 具体目标IV和V.制定快速收获HVT感染的 细胞，并从微载体中分离这些细胞， 高效价马立克氏病疫苗的生产 拟定商业应用： 生产了许多动物活疫苗（如马立克氏病疫苗）和人病毒疫苗 使用滚瓶系统或鸡胚中的鸡胚细胞 鸡蛋两种方法都没有使用微载体-生物反应器系统。大规模 在MiaoCarrier生物反应器系统中生产马立克氏疫苗，将导致 与旧技术相比，生物反应器具有显著的经济优势。 还应提高产品质量和一致性。和制造成本 降低本文提出的系统将利用（1）主或次CEF 细胞或永生化细胞系。(2)特殊培养基配方，（3）牛血清 并将发展（4）协议以在该系统中生产疫苗。的 这项技术应该推广到其他全细胞人类疫苗。 拟定商业应用： 目前人类流感疫苗供应短缺， 开发替代含胚蛋技术的重要性 生产方法。从（1）CDC对特定流感的估计到 病毒变种可能袭击美国，（2）疫苗的可用性大 规模非常长，许多个月。如果发生流感大流行 这种数月的准备时间可能导致高死亡率， 1917- 1918年大流行如果流感病毒疫苗能够大规模生产 生物反应器，周转时间快（几个月）， 一场大流行就可以避免。这项研究有可能解决 这个问题