Influenza neuraminidase active site proximity assay for rapid profiling of inhibitory antibodies and antigenic drift
Jin Gao,
Galina Landgraf,
Yue Yuan,
Hai Yu,
Soma Saeidi,
Hyeog Kang,
Mira Rakic Martinez,
Luca Giurgea,
Vladimir Lugovtsev,
Jason Gorman,
Matthew Memoli,
Xi Chen,
Zhiping Ye,
Robert Daniels
Affiliations
Jin Gao
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Galina Landgraf
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Yue Yuan
Department of Chemistry, University of California, One Shields Avenue
Hai Yu
Department of Chemistry, University of California, One Shields Avenue
Soma Saeidi
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Hyeog Kang
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Mira Rakic Martinez
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Luca Giurgea
Laboratory of Infectious Diseases Clinical Studies Unit, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Vladimir Lugovtsev
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Jason Gorman
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Matthew Memoli
Laboratory of Infectious Diseases Clinical Studies Unit, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health
Xi Chen
Department of Chemistry, University of California, One Shields Avenue
Zhiping Ye
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Robert Daniels
Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration
Abstract Efficient approaches that can help to select vaccine strains for the influenza virus neuraminidase (NA) antigen are currently needed to advance the development of vaccines containing NA. Here, we present a rapid and cost-effective solution-based NA active site proximity assay (NASPA) for measuring NA activity inhibitory (NAI) antibodies. This simplified assay uses large “bulky” NA active site-binding inhibitors to replace the sialylated glycoprotein substrates in common NA enzyme-linked lectin assay (ELLA) approaches. Our results with ferret antisera and monoclonal antibodies against vaccine strain NAs show a strong correlation between NASPA and ELLA titers, and that NASPA titers are not influenced by anti-HA antibodies. Consequently, NASPA can be used with influenza A or B strains and with the latter it revealed incremental antigenic changes in the NAs from recent B Victoria lineage vaccine strains. By coupling NASPA with a simple activity assay, we also found that steric and active site-binding NAI antibodies against circulating NAs are common in adult human sera. Finally, we demonstrate that NASPA can be modified by incorporating novel NA substrate-analog-based inhibitors. Together, these results suggest that NASPA can aid the development of vaccines containing NA by helping to select suitable vaccine strains and profile anti-NA antibody responses.
