A luciferase-based approach for measuring HBGA blockade antibody titers against human norovirus

Jessica M. van Loben Sels, Luke W. Meredith, Stanislav V. Sosnovtsev, Miranda de Graaf, Marion P.G. Koopmans, Lisa C. Lindesmith, Ralph S. Baric, Kim Y. Green*, Ian G. Goodfellow

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Background: Noroviruses are the most common cause of viral gastroenteritis worldwide, yet there is a deficit in the understanding of protective immunity. Surrogate neutralization assays have been widely used that measure the ability of antibodies to block virus-like particle (VLP) binding to histo-blood group antigens (HBGAs). However, screening large sample sets against multiple antigens using the traditional HBGA blocking assay requires significant investment in terms of time, equipment, and technical expertise, largely associated with the generation of purified VLPs. Methods: To address these issues, a luciferase immunoprecipitation system (LIPS) assay was modified to measure the norovirus-specific HBGA blockade activity of antibodies. The assay (designated LIPS-Blockade) was validated using a panel of well-characterized homotypic and heterotypic hyperimmune sera as well as strain-specific HBGA blocking monoclonal antibodies. Results: The LIPS-Blockade assay was comparable in specificity to a standard HBGA blocking protocol performed with VLPs. Using time-ordered patient sera, the luciferase-based approach was also able to detect changes in HBGA blocking titers following viral challenge and natural infection with norovirus. Conclusion: In this study we developed a rapid, robust, and scalable surrogate neutralization assay for noroviruses that circumvented the need for purified VLPs. This LIPS-Blockade assay should streamline the process of large-scale immunological studies, ultimately aiding in the characterization of protective immunity to human noroviruses.

Original languageEnglish
Article number114196
Number of pages11
JournalJournal of Virological Methods
Publication statusPublished - Nov 2021

Bibliographical note

Funding for this work was provided by the Wellcome Trust [207498/Z/17/Z] (IGG); the Division of Intramural Research of the National Institute of Allergy and Infectious Diseases, NIH (KYG); COMPARE grant agreement N°643476 (MPGK) and MRACE grant 2017 (MdG); and National Institute of Allergy and Infectious Diseases, NIH grants R01 AI148260, R56 AI106006, U19 AI109761 CETR, and the Wellcome Trust [203268/Z/16/Z] (RSB).

Publisher Copyright:
© 2021 Elsevier B.V.


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