A high-throughput multiplex assay to characterize flavivirus-specific immunoglobulins

Mélanie Merbah*, Suzanne Wollen-Roberts, Zhanna Shubin, Yifan Li, Hongjun Bai, Vincent Dussupt, Letzibeth Mendez-Rivera, Bonnie Slike, Shelly J. Krebs, Kayvon Modjarrad, Nelson L. Michael, Morgane Rolland

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Genus Flavivirus, which includes 53 virus species, is the leading cause of arthropod-borne diseases in humans. Diagnosis of these viral diseases is complicated by their overlapping epidemiology and clinical manifestations, and the fact that cross-reactive antibody responses are frequently elicited by individuals in response to infection. We developed a bead-based immunoassay to concomitantly profile the isotype and subclass of antibody responses (five isotypes and four subclasses) in parallel with specificity against multiple antigens. Our panel included 22 envelope (E) and non-structural 1 (NS1) proteins of different flaviviruses (Zika (ZIKV), Dengue (DENV), Yellow Fever (YFV), West Nile (WNV), Japanese Encephalitis (JEV) and Tick-Borne Encephalitis (TBEV)) and the envelope protein of Chikungunya virus (CHIKV). Using 54 samples from 40 individuals with ZIKV infection that had been pre-characterized, we identified 1) stronger ZIKV responses in individuals previously exposed to flavivirus compared to flavivirus-naïve individuals; 2) different antibody isotypes depending on the stage of infection: acute, convalescent and late convalescent; 3) cross-reactive responses; and 4) a potential CHIKV infection. The assay had a broad dynamic range (>5 logs) and has the potential to distinguish antigen-specific responses induced by ZIKV infection from cross-reactive responses. The multidimensional data provided by this high-throughput antibody-profiling platform can advance our understanding of the human immune response to flaviviruses as they expand their global reach.

Original languageEnglish
Article number112874
JournalJournal of Immunological Methods
StatePublished - Dec 2020
Externally publishedYes


  • Antibody detection
  • Cross-reactivity
  • Flavivirus
  • Immunoglobulin
  • ZIKV


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