Immunoassay for quantification of antigen-specific IgG fucosylation

Tonći Šuštić, Julie van Coillie, Mads Delbo Larsen, Ninotska I. L. Derksen, Zoltan Szittner, Jan Nouta, Wenjun Wang, Timon Damelang, Ianthe Rebergen, Federica Linty, Remco Visser, Juk Yee Mok, Dionne M. Geerdes, Wim J. E. van Esch, Steven W. de Taeye, Marit J. van Gils, Leo van de Watering, C. Ellen van der Schoot, Manfred Wuhrer, Theo RispensGestur Vidarsson

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2 Citations (Scopus)

Abstract

Background: Immunoglobulin G (IgG) antibodies serve a crucial immuno-protective function mediated by IgG Fc receptors (FcγR). Absence of fucose on the highly conserved N-linked glycan in the IgG Fc domain strongly enhances IgG binding and activation of myeloid and natural killer (NK) cell FcγRs. Although afucosylated IgG can provide increased protection (malaria and HIV), it also boosts immunopathologies in alloimmune diseases, COVID-19 and dengue fever. Quantifying IgG fucosylation currently requires sophisticated methods such as liquid chromatography-mass spectrometry (LC-MS) and extensive analytical skills reserved to highly specialized laboratories. Methods: Here, we introduce the Fucose-sensitive Enzyme-linked immunosorbent assay (ELISA) for Antigen-Specific IgG (FEASI), an immunoassay capable of simultaneously quantitating and qualitatively determining IgG responses. FEASI is a two-tier immunoassay; the first assay is used to quantify antigen-specific IgG (IgG ELISA), while the second gives FcγRIIIa binding-dependent readout which is highly sensitive to both the IgG quantity and the IgG Fc fucosylation (FcγR-IgG ELISA). Findings: IgG Fc fucosylation levels, independently determined by LC-MS and FEASI, in COVID-19 responses to the spike (S) antigen, correlated very strongly by simple linear regression (R2=0.93, p < 0.0001). The FEASI method was then used to quantify IgG levels and fucosylation in COVID-19 convalescent plasma which was independently validated by LC-MS. Interpretation: FEASI can be reliably implemented to measure relative and absolute IgG Fc fucosylation and quantify binding of antigen-specific IgG to FcγR in a high-throughput manner accessible to all diagnostic and research laboratories. Funding: This work was funded by the Stichting Sanquin Bloedvoorziening (PPOC 19-08 and SQI00041) and ZonMW 10430 01 201 0021.

Original languageEnglish
Article number104109
JournalEBIOMEDICINE
Volume81
DOIs
Publication statusPublished - 1 Jul 2022

Keywords

  • Anti-spike IgG
  • COVID-19
  • Fucosylation
  • IgG glycosylation

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