Monitoring System For Vector-Based Influenza Vaccine

描述(申请人提供)：这项先进技术建议书是对NIAID最近发布的高优先级流感研究领域的通知[NOT-AI-05-013]的回应。为了向卫生官员提供有效抗击流感大流行毒株所需的工具，至关重要的是开发快速、成本效益高的疫苗生产方法。病毒表达载体为以血凝素(HA)等抗原蛋白为基础的疫苗生产提供了一种有前途的策略。拟议的工作是为了支持开发疫苗生产的替代方法的努力。具体地说，拟议的工作重点是开发一个病毒和HA量化系统，该系统将显著改善过程控制，减少生产时间和成本，从而提高疫苗的生产能力。目的1.本研究的目标之一是开发一种创新的紧凑型双通道病毒计数器(DCVC)，并将其用于病毒表达载体系统的商业化应用。“完整”病毒数量代表“活跃”病毒的假设将得到严格检验。短期的重点将是蛋白质科学公司(PSC)为生产流感疫苗而开发的杆状病毒表达载体系统(BEVS)。然而，DCVC将同样适用于其他载体系统，例如GenVec目前正在开发的用于生产疟疾和艾滋病毒疫苗的腺载体技术。目标2.第二个目标是与DCVC一起开发一种分析方法，它将提供对BEVS疫苗生产过程中产生的HA的快速量化。要检验的假设是，DCVC可用于直接量化细胞表面的HA，该值与从细胞糊中分离出的水平相关。目前，PSC使用一种基于细胞活力的间接方法来评估细胞培养收获的HA，该方法与HA的产生相关。直接和快速地定量宿主细胞表面的HA将极大地加强杆状病毒表达系统的过程控制，并将显著降低疫苗的制造成本。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。 拟议的仪器和化验将有助于简化流感疫苗的生产。一旦发生流感大流行，快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.