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

芯片滴度：流感疫苗效力测定中 SRID 的替代方案

基本信息

批准号：
8465184
负责人：
KATHY L ROWLEN
金额：
$ 30万
依托单位：
INDEVR, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-15 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8465184
关键词：
Affinity Antibodies Antigens Binding Biological Assay Birds Calibration Development Evaluation Evaluation Research FDA approved Family suidae Fluorescence Glycoproteins Goals Gold Hand Hemagglutinin Hour Human Immunodiffusion Influenza Influenza Hemagglutinin Laboratories Manufacturer Name Measurement Measures Metric Monoclonal Antibodies National Institute of Allergy and Infectious Disease One-Step dentin bonding system Performance Pharmaceutical Preparations Preparation Printing Process Production Proteins Radial Reagent Recombinants Relative (related person)Research Sialic Acids Signal Transduction System Testing Time Training Vaccine Production Vaccine Research Vaccines Variant analytical method base cost cost effective density experience flu improved influenza virus vaccine interest response success vaccine development

项目摘要

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.

描述（由适用提供）：非常需要新的分析方法来增强疫苗的研发，最终允许在更少的时间内以更少的成本生产安全有效的疫苗。例如，众所周知，对于速率限制的过程中，SPLT疫苗的一个步骤是蛋白质数量和预算确定。 FDA批准的基于影响的血凝素蛋白疫苗的“金标准”电位测定是单辐射免疫接种（SRID）。 SRID是一个时间和劳动密集的测定法，通常需要2-3天才能完成，并且训练有素的分析师会准时至少6小时。尽管生物制剂评估与研究中心（CBER）无需提供参考试剂，但必须购买其他材料，并由每个疫苗生产商准备和验证整个测定法。疫苗生产商通常会在接收参考代理时经历长时间的延误。即使掌握了参考材料，在进行开发之前，每轮克隆评估的等待也可能是几天。众所周知，总体结果是耗时且无效的疫苗开发过程。在这里，我们提出了基于成本效益的低密度微阵列的两种新的定量，多重分析方法。这两种测定方法均基于CHIP？的滴度，该方法将通过大大减少结果的时间来简化疫苗效力测量，从而消除与测定准备相关的实验室间变化以及降低试剂成本。一种提出的分析依赖于（特定目的I）对造成影响的hemagglutinsuins types普遍反应的单克隆抗体（例如H1，H3，H5）。另一个提出的测定（特定目标II）依赖于连续的唾液酸糖蛋白结合血凝集素以实现快速HA蛋白的量，而无需菌株特异性抗体。我们认为，芯片上的滴度具有革新影响疫苗效力确定的长期潜力。