A Flexible High-Throughput Immunological Assay to Support Next-Generation Influenza Vaccine Studies

我们的目标是开发更好的检测流感感染后人体产生的抗体的方法。 或者接种疫苗。我们将开发测量总抗体反应的检测方法，并专门检测 测量负责预防感染的抗体。通过使用高吞吐量的多路传输格式 使用可以大批量制备和储存的试剂，分析将解决许多实用的 血凝抑制试验(HAI)和病毒微量试验等现行金标准方法的局限性 中和试验(MN)。检测将使用中尺度诊断®(MSD)电化学发光 (ECL)化验平台，已广泛用于新冠肺炎、肺炎球菌等的资格鉴定 疫苗。检测将使用多孔平板，每个孔中有流感血凝素(HA)阵列，以 支持同时测量针对疫苗中代表的毒株的抗体，并 分析针对其他感兴趣菌株的交叉保护。一套检测试剂和化验格式将是 当与抗原阵列组件一起使用时，它将支持一系列测量模式。 这些措施包括测量(I)阻断每个抗原识别唾液酸能力的抗体。 (2)抗每种抗原的总免疫球蛋白和/或免疫球蛋白A(在血液或呼吸道样本中)；和 (3)识别预先选定的通用表位的抗体。通过这项工作，我们将为 加快开发新型流感疫苗。 UH2/UH3分阶段计划将包括以下主要组成部分。在UH2可行性/早期 开发阶段我们将开发和优化检测抗体所需的检测试剂 对最近的10种疫苗毒株以及几种历史上的人类毒株和潜在的大流行的反应 禽类品系。在这一阶段，我们还将优化化验格式并组装样本集以供以后验证 工作。在UH3验证阶段，我们将建立生产检测试剂盒的制造程序，以 执行建议的检测方法，并通过测试特征良好的样本集和 将我们的结果与已建立的金标准方法进行比较。 †本建议书中标有星号的文件部分包含专有/机密信息，msd要求不得向其发布。 政府以外的人员，但审查和评估目的除外。

Our objective is to develop better assays for measuring antibodies produced in people after influenza infection or vaccination. We’ll develop assays that measure total antibody responses, and assays that specifically measure the antibodies responsible for protection from infection. By using high-throughput multiplexed formats employing reagents that can be prepared and stored in large lots, the assays will address many of the practical limitations of current gold standard methods like the hemagglutination inhibition assay (HAI) and the viral micro- neutralization (MN) assay. The assays will use the Meso Scale Diagnostics® (MSD) electrochemiluminescence (ECL) assay platform, which has been widely adopted for qualification of COVID-19, pneumococcal and other vaccines. The assays will use multi-well plates with arrays of influenza hemagglutinin (HA) in each well to support the simultaneous measurement of antibodies against the strains represented in a vaccine, and to analyze cross-protection against other strains of interest. A set of detection reagents and assay formats will be developed that, when used with the antigen array component, will support a range of measurement modalities. These include the measurement of (i) antibodies that block the ability of each antigen to recognize sialic acid (SA) modified host proteins; (ii) total IgG and/or IgA against each antigen (in blood or respiratory samples); and (iii) antibodies recognizing pre-selected universal epitopes. Through this work, we will provide important tools for accelerating the development of new influenza vaccines. The phased UH2/UH3 program will include the following main components. In the UH2 feasibility/early development phase we will develop and optimize the required assay reagents needed to measure antibody responses to 10 recent vaccine strains, as well as several historical human strains and potentially pandemic avian strains. In this phase, we will also optimize the assay formats and assemble sample sets for later validation work. In the UH3 validation phase, we will establish manufacturing procedures for producing assay kits to conduct the proposed assay methods, and validate performance by testing well-characterized sample sets and comparing our results to established gold standard methods. †The sections on documents contained in this proposal that are marked with an asterisk contain proprietary/confidential information that MSD requests not be released to persons outside the Government, except for purposes of review and evaluation.