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的研究和开发，最终允许在更短的时间内以更低的成本生产安全有效的疫苗。例如，众所周知，对于splt疫苗，过程中可能是速率限制的一个步骤是蛋白质定量和效价测定。FDA批准的基于流感血凝素蛋白的疫苗的“金标准”效力测定是单向放射免疫扩散（SRID）。SRID是一种时间和劳动密集型测定，通常需要2-3天才能完成，并且由训练有素的分析人员进行至少6小时的手工操作。虽然参考试剂由生物制品评价和研究中心（CBER）免费提供，但必须购买额外的材料，并由每个疫苗生产商制备和验证整个测定。疫苗生产商在收到参考试剂方面往往会经历长时间的拖延，有时长达数月。即使手头有参考材料，在向前发展之前，每一轮克隆评估等待结果的时间也可能是几天。正如人们广泛承认的那样，总体结果是疫苗开发过程耗时且效率低下。 在这里，我们提出了两个新的定量，多重分析方法的基础上，具有成本效益的低密度微阵列。两种检测方法均基于芯片上滴度方法，该方法将通过大幅缩短获得结果的时间、消除与检测制备相关的实验室间差异以及降低试剂成本来简化疫苗效价测量。一种提出的测定依赖于（特异性目标I）对流感的血凝素亚型普遍应答的单克隆抗体（例如，H1、H3、H5）。另一种拟定测定（特异性目的II）依赖于结合血凝素的通用唾液酸糖蛋白，以实现快速HA蛋白定量，而无需菌株特异性抗体。我们相信，芯片滴度具有彻底改变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.