A MICROCARRIER/CEF/MEDIA SYSTEM TO MAKE VIRAL VACCINES

这项研究旨在开发一种在生物反应器中的微载体上培养鸡胚胎成纤维细胞(CEF)的系统，用于大规模生产禽流感和人类病毒疫苗。滚筒瓶中的CEF细胞被用于生产马立克和纽卡斯尔等禽类疫苗。胚胎卵子(CEF)被用来制造流感等人类疫苗。由于这两个生产过程都是劳动密集型的，并且容易受到工艺变化的影响，因此将这些过程转换到受控的生物反应器环境将产生更一致和更低成本的产品。对鸡蛋制品过敏的疫苗接受者将会松一口气。具体目的是为了展示我们最新的两种微载体衬底的可行性，正如初步数据所显示的那样，这两种衬底具有很好的前景。具体目的一.检测(1)生长在ProNectin F包被的微载体上的原代CEF细胞和(2)用于生产火鸡疱疹病毒(马立克氏病)的三甲胺核心微载体，并比较(3)用微载体培养的细胞与用滚瓶和/或胚胎鸡蛋培养的细胞获得的病毒产量。具体目的II和III。使用(1)二次传代CEF细胞，(2)建立的CEF细胞系和(3)定制配方的培养液进行实验。拟议的商业应用：目前，原代鸡胚胎成纤维细胞(CEF)有巨大且众多的工业应用。动物疫苗(如马立克氏疫苗)和人类疫苗(如流感疫苗)都是使用CEF细胞生产的，一些工艺使用滚瓶中的CEF，一些工艺使用胚胎鸡蛋中的CEF，但没有一个使用生物反应器中微载体上的CEF。通过将CEF细胞应用于生物反应器中的微载体，能够减少劳动力含量，这对所有疫苗制造商都是经济上有利的。我们提出的这一新系统将包括CEF细胞、微载体和定制介质。

The research proposed is aimed at developing a system for growing chick embryo fibroblast (CEF) cells on microcarriers in bioreactors for the large- scale production of avian and human virus vaccines. CEF cells in roller bottles are used to manufacture avian vaccines such as Marek's and Newcastle. Embryonated eggs (CEF) are used to manufacture human vaccines such as influenza. Since both manufacturing processes are labor intensive and subject to process variations, conversion of these processes to a controlled bioreactor environment would result in more consistent and lower cost products. Vaccine recipients who are allergic to egg products will be relieved. The Specific Aims are directed at showing the feasibility with two of our newest microcarrier substrates, which have promise as the preliminary data show. Specific Aim I. To examine (1) primary CEF cells grown on ProNectin F coated microcarriers and (2) trimethylamine core microcarriers for production of the herpesvirus of turkeys (Marek's disease) and compare (3) virus yields obtained with microcarrier-grown cells to yields obtained with cells grown in roller bottles and/or in embryonated eggs. Specific Aim II and III. To conduct experiments utilizing (1) secondary passage CEF cells, (2) an established CEF cell line and (3) custom formulated media. PROPOSED COMMERCIAL APPLICATIONS: At the present time there are huge, and numerous, industrial applications for primary chick embryo fibroblast (CEF) cells. Both animal vaccines (e.g. Marek's) and human vaccines (e.g. influenza) are manufactured using CEF cells, some processes use CEFs in roller bottles, some are in embryonated eggs, but none use CEFs on microcarriers in bioreactors. It would be economically advantageous to all the vaccine manufacturers to be able to reduce labor content by applying CEF cells to microcarriers in bioreactors. This new system we propose, will include CEF cells, microcarriers and the custom media.