Monitoring System For Vector-Based Influenza Vaccine

描述（由申请人提供）：本先进技术提案是对NIAID最近发布的高优先级流感研究领域通知的回应[NOT-AI-05-013]。为了向卫生官员提供有效防治流感大流行株所需的工具，必须开发快速和具有成本效益的疫苗生产方法。病毒表达载体为基于抗原蛋白如血凝素（HA）的疫苗的生产提供了有前景的策略。拟议的工作是为了支持开发疫苗生产替代方法的努力。具体而言，拟议的工作集中在病毒和HA定量系统的开发上，该系统将显著改善过程控制并减少生产时间和成本，从而提高疫苗生产能力。目的1.该研究的一个目标是开发一种创新的紧凑型双通道病毒计数器（DCVC）和用于病毒表达载体系统的商业应用的检测方法。将严格检验“完整”病毒计数代表“活性”病毒的假设。短期重点将放在由Protein Sciences Corporation（PSC）开发的用于生产流感疫苗的杆状病毒表达载体系统（BEVS）上。然而，DCVC同样适用于其他载体系统，例如目前正在开发用于生产疟疾和HIV疫苗的GenVec腺病毒载体技术。目标2.第二个目标是开发一种与DCVC结合的检测方法，该方法将提供BEVS疫苗生产过程中产生的HA的快速定量。待检验的假设是DCVC可用于直接定量细胞表面上的HA，并且该值与从细胞糊中分离的水平相关。目前，PSC使用基于细胞活力的间接方法评估HA的细胞培养收获物，细胞活力与HA产量相关。直接和快速定量宿主细胞表面上的HA将大大提高杆状病毒表达系统的过程控制，并将导致疫苗生产成本的显着降低。监测系统的这两个目标都需要FDA批准才能应用于疫苗生产。 拟议的仪器和分析将有助于简化流感疫苗的生产。如果发生流感大流行，迅速生产疫苗将在最大限度地减少生命损失方面发挥关键作用。

DESCRIPTION (provided by applicant): This Advanced Technology proposal is in response to NIAID's recently issued Notice of High-Priority Influenza Research Areas [NOT-AI-05-013]. In order to provide health officials with the tools required to efficiently combat a pandemic strain of influenza, it is essential that rapid and cost-effective methods for vaccine production be developed. Viral expression vectors offer a promising strategy for production of vaccines based on antigenic proteins such as hemagglutinin (HA). The proposed work is in support of efforts to develop alternate methods for vaccine production. Specifically, the proposed work centers on the development of a virus and HA quantification system that would significantly improve process control and reduce production time and costs, thereby enhancing vaccine production capabilities. Objective 1. One goal of the proposed research is to develop an innovative compact dual channel virus counter (DCVC) and assay for commercial application to viral expression vector systems. The hypothesis that a count of "intact" viruses is representative of "active" viruses will be rigorously tested. The short-term focus will be on the baculovirus expression vector system (BEVS) developed by Protein Sciences Corporation (PSC) for the generation of influenza vaccine. However, the DCVC would be equally applicable to other vector systems, such as the GenVec's adenovector technology currently being developed for production of malaria and HIV vaccines. Objective 2. The second goal is to develop an assay, in conjunction with the DCVC, which will provide rapid quantification of HA generated during the BEVS vaccine production process. The hypothesis to be tested is that the DCVC can be used to directly quantify HA on cell surfaces and that value is correlated with levels isolated from cell paste. Currently, PSC evaluates cell culture harvest of HA using an indirect method based on cell viability, which is correlated with HA production. Direct and rapid quantification of HA on the host cell surface during would greatly enhance process control for the baculovirus expression system and would result in significant reduction in manufacturing cost for vaccines. Both objectives for the monitoring system will require FDA approval for application to vaccine production. The proposed instrument and assays would aid in streamlining influenza vaccine production. In the event of an influenza pandemic, rapid vaccine production will play a key role in minimizing loss of life.