TITER ON CHIP: ALTERNATIVE TO SRID FOR INFLUENZA VACCINE POTENCY DETERMINATION

描述(由申请人提供)：迫切需要新的分析方法来加强疫苗研究和开发，最终以更低的成本在更短的时间内生产安全有效的疫苗。例如，众所周知，对SPLT疫苗来说，限速过程中的一个步骤是蛋白质定量和效力测定。FDA批准的流感血凝素蛋白疫苗效价的金标准是单径向免疫扩散法(SRID)。SRID是一项耗时耗力的分析，通常需要2-3天的时间才能完成，并由训练有素的分析师至少6小时亲自动手。虽然参考试剂由生物制品评估和研究中心(CBER)免费提供，但必须购买额外的材料，并由每个疫苗生产商准备和验证整个分析。疫苗生产商在收到参考试剂方面往往会经历很长时间的延误，有时甚至是几个月。即使有了参考材料，在推进开发之前，每一轮克隆评估的结果也可能需要几天的等待。正如广泛承认的那样，总体结果是疫苗开发过程既耗时又低效。在这里，我们提出了两种新的基于成本效益低密度微阵列的定量、多路分析方法。这两种检测方法都是基于芯片滴度方法，该方法将通过大幅缩短得出结果的时间、消除与分析准备相关的实验室间差异以及降低试剂成本来简化疫苗效力测量。一种建议的检测依赖于(特定目标I)对流感血凝素亚型(例如，H1、H3、H5)普遍有反应的单抗。另一种被提议的检测方法(特异性目标II)依赖于结合血凝素的通用唾液酸糖蛋白来实现快速的HA蛋白定量，而不需要菌株特异性抗体。我们相信，芯片上的滴度具有革命性的流感疫苗效力测定的长期潜力。 与公共卫生相关：迫切需要新的分析方法来加强疫苗研究和开发，最终以更低的成本在更短的时间内生产安全有效的疫苗。在这里，我们提出了两种新的基于成本效益低密度微阵列的定量、多路分析方法。我们相信，芯片上的滴度具有革命性的流感疫苗效力测定的长期潜力。

DESCRIPTION (provided by applicant): There is a tremendous need for new analytical methods to enhance vaccine research and development, ultimately allowing the production of safe and efficacious vaccines in less time at less cost. For example, it is well known that for splt vaccines one step in the process that can be rate limiting is protein quantification and potency determination. The FDA approved "gold standard" potency assay for influenza hemagglutinin protein based vaccines is single radial immunodiffusion (SRID). SRID is a time and labor intensive assay, often requiring 2-3 days to complete and a minimum of 6 hours hands on time by well trained analysts. While the reference reagents are provided at no cost by the Center for Biologics Evaluation and Research (CBER), additional materials must be purchased and the entire assay prepared and validated by each vaccine producer. Often vaccine producers experience long delays, sometime months, in receiving reference reagents. Even with reference materials in hand, the wait for results can be days for each round of clone assessment prior to moving forward in development. As is widely acknowledged, the overall result is a time-consuming and inefficient vaccine development process. Here we propose two new quantitative, multiplexed analytical methods based on cost-effective low density microarrays. Both assays are based on a ¿Titer on Chip¿ approach that will streamline vaccine potency measurements by substantially reducing time to result, eliminating inter-laboratory variations associated with assay preparations, and reducing reagent cost. One proposed assay relies (Specific Aim I) on monoclonal antibodies that are universally responsive to hemagglutinin subtypes for influenza (e.g., H1, H3, H5). The other proposed assay (Specific Aim II) relies on universal sialic acid glycoproteins that bind hemagglutinin to achieve rapid HA protein quantification without the need for strain specific antibodies. We believe that Titer on Chip has the long term potential to revolutionize influenza vaccine potency determination. PUBLIC HEALTH RELEVANCE: There is a tremendous need for new analytical methods to enhance vaccine research and development, ultimately allowing the production of safe and efficacious vaccines in less time at less cost. Here we propose two new quantitative, multiplexed analytical methods based on cost-effective low density microarrays. We believe that Titer on Chip has the long term potential to revolutionize influenza vaccine potency determination.