岩藻糖基化
免疫分析
免疫球蛋白G
抗原
抗体
免疫学
化学
聚糖
分子生物学
病毒学
生物
糖蛋白
作者
Tonći Šuštić,Julie Van Coillie,Mads Delbo Larsen,Ninotska I. L. Derksen,Zoltán 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 Rispens,Gestur Vidarsson
出处
期刊:EBioMedicine
[Elsevier]
日期:2022-07-01
卷期号:81: 104109-104109
被引量:6
标识
DOI:10.1016/j.ebiom.2022.104109
摘要
BackgroundImmunoglobulin 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.MethodsHere, 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).FindingsIgG 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.InterpretationFEASI 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.FundingThis work was funded by the Stichting Sanquin Bloedvoorziening (PPOC 19-08 and SQI00041) and ZonMW 10430 01 201 0021.
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