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方法来测量流感感染后人体产生的抗体 或接种疫苗。我们将开发测量总抗体反应的检测方法， 测量负责保护免受感染的抗体。通过使用高通量多路复用格式 使用可以大量制备和储存的试剂，该测定将解决许多实际问题。 目前金标准方法的局限性，如血凝抑制试验（HAI）和病毒微量 中和（MN）测定。检测将使用Meso Scale Diagnostics®（MSD）电化学发光 (ECL)检测平台，已被广泛用于COVID-19、肺炎球菌和其他 疫苗。该检测将使用多孔板，每个孔中含有流感血凝素（HA）阵列， 支持同时测量针对疫苗中代表的菌株的抗体，以及 分析针对其他感兴趣菌株的交叉保护。将提供一套检测试剂和检测格式， 开发的，当与抗原阵列组件一起使用时，将支持一系列测量模式。 这些包括测量（i）阻断每种抗原识别唾液酸的能力的抗体 (SA)（ii）针对每种抗原的总IgG和/或伊加（在血液或呼吸道样品中）;和 (iii)识别预选通用表位的抗体。通过这项工作，我们将提供重要的工具， 加快研制新型流感疫苗。 UH 2/UH 3分阶段计划将包括以下主要组成部分。在UH 2可行性/早期 在开发阶段，我们将开发和优化所需的检测试剂， 对10种最近的疫苗株以及几种历史上的人类毒株和潜在的大流行毒株的反应 鸟类菌株。在此阶段，我们还将优化分析格式并组装样本集以供后续验证 工作在UH 3验证阶段，我们将建立生产检测试剂盒的生产程序， 执行拟定的测定方法，并通过检测充分表征的样品集来验证性能， 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默沙东要求不得向其披露的专有/机密信息。 政府以外的人，但为了审查和评估的目的除外。

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.