University of Minnesota
https://twin-cities.umn.edu/
612-625-5000
Milestone
2.1.b

Reference reagents and assay protocols

In progress

Develop standardized reference reagents and harmonized assay protocols for studying immunoglobulin responses to influenza infection and vaccination, such as qualified assays for detecting mucosal antibodies.

Progress Highlights

Waldock 2023 (FLUCOP consortium) conducted a head-to-head comparison of harmonized HAI and MN protocols to evaluate the relationship between HAI and MN titers, and the impact of assay harmonization and standardization on inter-laboratory variability, and agreement between these methods.

See research
 

Waldock 2023 (FLUCOP consortium) developed a feasibility study for conducting an external quality assessment scheme for influenza serology methods, to improve performance, reduce inter-laboratory variation, and support the use of harmonized protocols and biological standards for seasonal influenza serology testing.

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Bernard 2022 (FLUCOP Consortium) present a freely available and detailed standard operating procedure for N1 influenza antigen using a validated ELLA-NI. The assay performed consistently with both influenza A and influenza B antigens. 

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Waldock 2021 (FLUCOP consortium) assessed the impact of harmonizing the HAI assay protocol/reagents and using standards on interlaboratory variability. 

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NIAID CIVICs is qualifying assays including secretory IgA MSD to look at mucosal response to novel flu vaccines; Duke, University of Maryland—Baltimore, and Imperial College London have harmonized protocols and are conducting proficiency panels to measure harmonization; reagents, viruses, mAb are available at these sites or in the BEI Resources portal. NIAID has also identified mAb panels for use in assays for CIVICs trials.

See BEI Resources portal

Vincent 2024 developed a high-throughput novel modified validated HAI assay for evaluating vaccine immunogenicity and efficacy using egg-derived influenza viruses or recombinant VLPs as agglutinins and human RBCs to measure anti-influenza HA Ab titers in human serum, to test sera from influenza vaccine clinical trials. Results showed that titers from the validated HAI assay correlated well with those of a qualified microneutralization assay.

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Koutsakos 2023 assessed the utility of different types of samples from the human respiratory tract, including nasopharyngeal swabs (for assessing antibodies in nasal mucosa) and bronchoalveolar lavage (for assessing antibodies from the lower respiratory tract). This study informs the value of different sample types for measuring mucosal Abs.

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Levy 2023 developed a novel antigen microarray-based assay for rapid high-throughput antibody profiling, which can be used for profiling IgG, IgA, and IgM responses to multiple antigens simultaneously, requiring minimal amounts of sample and antigens.

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Rumfelt 2023 developed a luciferase MN assay that combines the advantages of the conventional MN assay with the ease of the HAI assay.

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Sawant 2023 developed a qualified and adaptable influenza serology method and analysis strategy to measure quantifiable HAI titers to define correlates of vaccine-mediated protection in human clinical trials. 

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NIAID CIVICs is qualifying assays including secretory IgA MSD to look at mucosal response to novel flu vaccines; Duke, University of Maryland—Baltimore, and Imperial College London have harmonized protocols and are conducting proficiency panels to measure harmonization; reagents, viruses, mAb are available at these sites or in the BEI Resources portal. NIAID has also identified mAb panels for use in assays for CIVICs trials.

See BEI Resources portal