Immunoassay for quantification of antigen-specific IgG fucosylation

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

doi:10.1016/j.ebiom.2022.104109

Cited by 10 publications

(13 citation statements)

References 62 publications

(137 reference statements)

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“…Despite the low abundance and relatively stable concentrations of afucosylated IgG in plasma, multiple studies of antigen-specific IgG, all using affinity-purified IgG and mass spectrometry analysis, recently highlighted the clinical importance of some antigen-specific afucosylated IgG responses, which were also verified by FcγRIIIa-dependent serological methods in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) responses [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][63][64][65]. These afucosylated IgG responses include those directed towards alloantigens (Table 1), which can contribute to alloimmune-mediated diseases (Box 1), against enveloped viruses, and against P. falciparum proteins expressed on red blood cells (RBCs), all responses in which the degree of afucosylation can influence disease severity [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][63][64][65].…”

Section: Afucosylated Igg Responses In Alloimmunity and Infectious Di...mentioning

confidence: 99%

Afucosylated IgG responses in humans – structural clues to the regulation of humoral immunity

Oosterhoff

Larsen

Schoot

et al. 2022

Trends in Immunology

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Section: Afucosylated Igg Responses In Alloimmunity and Infectious Di...mentioning

confidence: 99%

Afucosylated IgG responses in humans – structural clues to the regulation of humoral immunity

Oosterhoff

Larsen

Schoot

et al. 2022

Trends in Immunology

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“…This method utilizes the enhanced affinity of FcγRIIIa for antigen-specific IgG fucosylation and translates the in vivo affinity and avidity enhancement into functional receptor binding properties. 58 The readout of this functional assay correlated directly with fucosylation values obtained by MS, linking the MSobtained antibody glyco-profiling with functional differences in FcγRIIIa binding. 58 Pro-inflammatory responses of afucosylated IgG are seen in FcγRIIImediated pathologies in patients with severe dengue fever 24 and alloimmunity.…”

Section: Discussionmentioning

confidence: 89%

“…58 The readout of this functional assay correlated directly with fucosylation values obtained by MS, linking the MSobtained antibody glyco-profiling with functional differences in FcγRIIIa binding. 58 Pro-inflammatory responses of afucosylated IgG are seen in FcγRIIImediated pathologies in patients with severe dengue fever 24 and alloimmunity. 20 Afucosylated antibody levels furthermore correlated with disease severity of COVID-19 patients.…”

Section: Discussionmentioning

confidence: 89%

The BNT162b2 mRNA SARS-CoV-2 Vaccine Induces Transient Afucosylated IgG1 in Naive But Not in Antigen-Experienced Vaccinees

Coillie

Pongrácz²,

Rahmöller³

et al. 2022

SSRN Journal

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“…Taken together, our results confirm that FcγRs are in many ways superior to anti-IgG antibodies, not only because they unravel the biological response but also because their application enable a lower background on cases where the investigated cells themselves possess FcγRs (such as platelet FcγRIIa), and therefore may carry cytophilic antibodies.. The potential benefits of this approach using recombinant FcγRs have already been appreciated in recent works from multiple groups ( 64 – 66 ), in particular the use of the FcγRIIIa receptor to characterize IgG fucosylation ( 67 ). Here, we have expanded this repertoire to include the detection and characterization of platelet-specific allo- and autoantibodies.…”

Section: Discussionmentioning

confidence: 99%

Cellular surface plasmon resonance-based detection of anti-HPA-1a antibody glycosylation in fetal and neonatal alloimmune thrombocytopenia

Szittner,

Bentlage,

Temming

et al. 2023

Front. Immunol.

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Fetal and neonatal alloimmune thrombocytopenia (FNAIT) can occur due to maternal IgG antibodies targeting platelet antigens, causing life-threatening bleeding in the neonate. However, the disease manifests itself in only a fraction of pregnancies, most commonly with anti-HPA-1a antibodies. We found that in particular, the core fucosylation in the IgG-Fc tail is highly variable in anti-HPA-1a IgG, which strongly influences the binding to leukocyte IgG-Fc receptors IIIa/b (FcγRIIIa/b). Currently, gold-standard IgG-glycoanalytics rely on complicated methods (e.g., mass spectrometry (MS)) that are not suited for diagnostic purposes. Our aim was to provide a simplified method to quantify the biological activity of IgG antibodies targeting cells. We developed a cellular surface plasmon resonance imaging (cSPRi) technique based on FcγRIII-binding to IgG-opsonized cells and compared the results with MS. The strength of platelet binding to FcγR was monitored under flow using both WT FcγRIIIa (sensitive to Fc glycosylation status) and mutant FcγRIIIa-N162A (insensitive to Fc glycosylation status). The quality of the anti-HPA-1a glycosylation was monitored as the ratio of binding signals from the WT versus FcγRIIIa-N162A, using glycoengineered recombinant anti-platelet HPA-1a as a standard. The method was validated with 143 plasma samples with anti-HPA-1a antibodies analyzed by MS with known clinical outcomes and tested for validation of the method. The ratio of patient signal from the WT versus FcγRIIIa-N162A correlated with the fucosylation of the HPA-1a antibodies measured by MS (r=-0.52). Significantly, FNAIT disease severity based on Buchanan bleeding score was similarly discriminated against by MS and cSPRi. In conclusion, the use of IgG receptors, in this case, FcγRIIIa, on SPR chips can yield quantitative and qualitative information on platelet-bound anti-HPA-1a antibodies. Using opsonized cells in this manner circumvents the need for purification of specific antibodies and laborious MS analysis to obtain qualitative antibody traits such as IgG fucosylation, for which no clinical test is currently available.

show abstract

Immunoassay for quantification of antigen-specific IgG fucosylation

Cited by 10 publications

References 62 publications

Afucosylated IgG responses in humans – structural clues to the regulation of humoral immunity

Afucosylated IgG responses in humans – structural clues to the regulation of humoral immunity

The BNT162b2 mRNA SARS-CoV-2 Vaccine Induces Transient Afucosylated IgG1 in Naive But Not in Antigen-Experienced Vaccinees

Cellular surface plasmon resonance-based detection of anti-HPA-1a antibody glycosylation in fetal and neonatal alloimmune thrombocytopenia

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