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Immunoassay for quantification of antigen-specific IgG fucosylation.
Sustic, Tonci; Van Coillie, Julie; Larsen, Mads Delbo; Derksen, Ninotska I L; Szittner, Zoltan; Nouta, Jan; Wang, Wenjun; Damelang, Timon; Rebergen, Ianthe; Linty, Federica; Visser, Remco; Mok, Juk Yee; Geerdes, Dionne M; van Esch, Wim J E; de Taeye, Steven W; van Gils, Marit J; van de Watering, Leo; van der Schoot, C Ellen; Wuhrer, Manfred; Rispens, Theo; Vidarsson, Gestur.
  • Sustic T; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Van Coillie J; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Larsen MD; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Derksen NIL; Department of Immunopathology, Sanquin Research, Amsterdam, the Netherlands; Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands.
  • Szittner Z; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Nouta J; Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands.
  • Wang W; Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands.
  • Damelang T; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Rebergen I; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Linty F; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Visser R; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Mok JY; Sanquin Reagents, Amsterdam, the Netherlands.
  • Geerdes DM; Sanquin Reagents, Amsterdam, the Netherlands.
  • van Esch WJE; Sanquin Reagents, Amsterdam, the Netherlands.
  • de Taeye SW; Department of Medical Microbiology, Amsterdam UMC, Amsterdam Infection and Immunity Institute, University of Amsterdam, Amsterdam, the Netherlands.
  • van Gils MJ; Department of Medical Microbiology, Amsterdam UMC, Amsterdam Infection and Immunity Institute, University of Amsterdam, Amsterdam, the Netherlands.
  • van de Watering L; Unit for Transfusion Medicine, Blood Bank, Sanquin, Amsterdam, the Netherlands.
  • van der Schoot CE; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
  • Wuhrer M; Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands.
  • Rispens T; Department of Immunopathology, Sanquin Research, Amsterdam, the Netherlands; Landsteiner Laboratory, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands.
  • Vidarsson G; Department of Experimental Immunohematology, Sanquin Research, Amsterdam, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands. Electronic
EBioMedicine ; 81: 104109, 2022 Jul.
Article in English | MEDLINE | ID: covidwho-1906947
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.
Subject(s)
Keywords

Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoglobulin G / Receptors, IgG / Fucose Type of study: Diagnostic study / Prognostic study / Qualitative research Limits: Humans Language: English Journal: EBioMedicine Year: 2022 Document Type: Article Affiliation country: J.ebiom.2022.104109

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoglobulin G / Receptors, IgG / Fucose Type of study: Diagnostic study / Prognostic study / Qualitative research Limits: Humans Language: English Journal: EBioMedicine Year: 2022 Document Type: Article Affiliation country: J.ebiom.2022.104109