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

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.