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  1. Article ; Online: Functional blood cell analysis by label-free biosensors and single-cell technologies.

    Szittner, Zoltán / Péter, Beatrix / Kurunczi, Sándor / Székács, Inna / Horvath, Robert

    Advances in colloid and interface science

    2022  Volume 308, Page(s) 102727

    Abstract: In this review we aim to summarize the current state of methods for label-free identification and functional characterization of leukocytes with biosensors and novel single cell techniques. The growing interest in this field is fueled from multiple ... ...

    Abstract In this review we aim to summarize the current state of methods for label-free identification and functional characterization of leukocytes with biosensors and novel single cell techniques. The growing interest in this field is fueled from multiple directions, with the different aspects highlighting benefits of these novel technologies in comparison to classical methods. The advantage of label-free characterization is that labeling the cells might affect their behavior, and therefore lead to a biased description of the investigated biological phenomena. Label-free biosensors can offer the benefit of (i) decreasing processing time and reagent costs, (ii) enable point-of-care diagnostics, and (iii) allow downstream application of the investigated cells. Moreover, (iv) label-free detection allows the monitoring of real-time kinetic processes, opening up new avenues in contrast to traditional structural characterizations. The emphasis in the review will be on techniques on the characterizations of single cells with special attention to surface sensitive technologies. Recent developments highlighted the importance of small cell populations and individual cells both in health and disease. Nonetheless techniques capable of analyzing single cells offer a promising tool for therapeutic approaches where characterization of individual cells is necessary to estimate their clinical therapeutic potential. Most of the approaches discussed here will cover the cellular activation, adhesion as measured on functionalized solid substrates, since this approach offers the most advantages. Analyzing various cells on solid substrates not only allows their individual morphological characterization and therefore a more precise description of their activation, but as well offers an opportunity to design multiplex measurements. With this approach different stimuli can be investigated in parallel and measure cellular avidity to targets, an important aspect of gaining more and more attention recently in characterization of T-cells and antibody effector functions. Finally, novel label-free approaches provide a solution to extracting unlabeled cells for downstream processing (e.g., transcriptome analysis, cloning or the aforementioned clinical potential), where ongoing and potential further applications are discussed.
    MeSH term(s) Biosensing Techniques/methods ; Blood Cells
    Language English
    Publishing date 2022-07-06
    Publishing country Netherlands
    Document type Journal Article ; Review
    ZDB-ID 210507-x
    ISSN 1873-3727 ; 0001-8686
    ISSN (online) 1873-3727
    ISSN 0001-8686
    DOI 10.1016/j.cis.2022.102727
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Polysaccharide-based nano-engineered multilayers for controlled cellular adhesion in label-free biosensors.

    Wasilewska, Monika / Michna, Aneta / Pomorska, Agata / Wolski, Karol / Zapotoczny, Szczepan / Farkas, Enikő / Szittner, Zoltan / Szekacs, Inna / Horvath, Robert

    International journal of biological macromolecules

    2023  Volume 247, Page(s) 125701

    Abstract: Controlling cellular adhesion is a critical step in the development of biomaterials, and in cell- based biosensing assays. Usually, the adhesivity of cells is tuned by an appropriate biocompatible layer. Here, synthetic poly(diallyldimethylammonium ... ...

    Abstract Controlling cellular adhesion is a critical step in the development of biomaterials, and in cell- based biosensing assays. Usually, the adhesivity of cells is tuned by an appropriate biocompatible layer. Here, synthetic poly(diallyldimethylammonium chloride) (PDADMAC), natural chitosan, and heparin (existing in an extracellular matrix) were selected to assembly PDADMAC/heparin and chitosan/heparin films. The physicochemical properties of macroion multilayers were determined by streaming potential measurements (SPM), quartz crystal microbalance (QCM-D), and optical waveguide lightmode spectroscopy (OWLS). The topography of the wet films was imaged using atomic force microscopy (AFM). The adhesion of preosteoblastic cell line MC3T3-E1 on those well-characterized polysaccharide-based multilayers was evaluated using a resonant waveguide grating (RWG) based optical biosensor and digital holographic microscopy. The latter method was engaged to investigate long-term cellular behavior on the fabricated multilayers. (PDADMAC/heparin) films were proved to be the most effective in inducing cellular adhesion. The cell attachment to chitosan/heparin-based multilayers was negligible. It was found that efficient adhesion of the cells occurs onto homogeneous and rigid multilayers (PDADMAC/heparin), whereas the macroion films forming "sponge-like" structures (chitosan/heparin) are less effective, and could be employed when reduced adhesion is needed. Polysaccharide-based multilayers can be considered versatile systems for medical applications. One can postulate that the presented results are relevant not only for modeling studies but also for applied research.
    MeSH term(s) Chitosan/chemistry ; Polysaccharides/pharmacology ; Heparin/pharmacology ; Heparin/chemistry ; Biosensing Techniques ; Cell Adhesion ; Surface Properties
    Chemical Substances Chitosan (9012-76-4) ; poly-N,N-dimethyl-N,N-diallylammonium chloride (26062-79-3) ; Polysaccharides ; Heparin (9005-49-6)
    Language English
    Publishing date 2023-07-08
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 282732-3
    ISSN 1879-0003 ; 0141-8130
    ISSN (online) 1879-0003
    ISSN 0141-8130
    DOI 10.1016/j.ijbiomac.2023.125701
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  3. Article ; Online: Review of Label-Free Monitoring of Bacteria: From Challenging Practical Applications to Basic Research Perspectives.

    Péter, Beatrix / Farkas, Eniko / Kurunczi, Sandor / Szittner, Zoltán / Bősze, Szilvia / Ramsden, Jeremy J / Szekacs, Inna / Horvath, Robert

    Biosensors

    2022  Volume 12, Issue 4

    Abstract: Novel biosensors already provide a fast way to detect the adhesion of whole bacteria (or parts of them), biofilm formation, and the effect of antibiotics. Moreover, the detection sensitivities of recent sensor technologies are large enough to investigate ...

    Abstract Novel biosensors already provide a fast way to detect the adhesion of whole bacteria (or parts of them), biofilm formation, and the effect of antibiotics. Moreover, the detection sensitivities of recent sensor technologies are large enough to investigate molecular-scale biological processes. Usually, these measurements can be performed in real time without using labeling. Despite these excellent capabilities summarized in the present work, the application of novel, label-free sensor technologies in basic biological research is still rare; the literature is dominated by heuristic work, mostly monitoring the presence and amount of a given analyte. The aims of this review are (i) to give an overview of the present status of label-free biosensors in bacteria monitoring, and (ii) to summarize potential novel directions with biological relevancies to initiate future development. Optical, mechanical, and electrical sensing technologies are all discussed with their detailed capabilities in bacteria monitoring. In order to review potential future applications of the outlined techniques in bacteria research, we summarize the most important kinetic processes relevant to the adhesion and survival of bacterial cells. These processes are potential targets of kinetic investigations employing modern label-free technologies in order to reveal new fundamental aspects. Resistance to antibacterials and to other antimicrobial agents, the most important biological mechanisms in bacterial adhesion and strategies to control adhesion, as well as bacteria-mammalian host cell interactions are all discussed with key relevancies to the future development and applications of biosensors.
    MeSH term(s) Anti-Bacterial Agents ; Anti-Infective Agents ; Bacteria ; Biosensing Techniques/methods
    Chemical Substances Anti-Bacterial Agents ; Anti-Infective Agents
    Language English
    Publishing date 2022-03-22
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2662125-3
    ISSN 2079-6374 ; 2079-6374
    ISSN (online) 2079-6374
    ISSN 2079-6374
    DOI 10.3390/bios12040188
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Cellular surface plasmon resonance-based detection of anti-HPA-1a antibody glycosylation in fetal and neonatal alloimmune thrombocytopenia.

    Szittner, Zoltán / Bentlage, Arthur E H / Temming, A Robin / Schmidt, David E / Visser, Remco / Lissenberg-Thunnissen, Suzanne / Mok, Juk Yee / van Esch, Wim J E / Sonneveld, Myrthe E / de Graaf, Erik L / Wuhrer, Manfred / Porcelijn, Leendert / de Haas, Masja / van der Schoot, C Ellen / Vidarsson, Gestur

    Frontiers in immunology

    2023  Volume 14, Page(s) 1225603

    Abstract: 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 ... ...

    Abstract 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.
    MeSH term(s) Pregnancy ; Female ; Infant, Newborn ; Humans ; Thrombocytopenia, Neonatal Alloimmune/diagnosis ; Surface Plasmon Resonance/methods ; Glycosylation ; Blood Platelets ; Immunoglobulin G ; Hemorrhage
    Chemical Substances Immunoglobulin G
    Language English
    Publishing date 2023-10-05
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2606827-8
    ISSN 1664-3224 ; 1664-3224
    ISSN (online) 1664-3224
    ISSN 1664-3224
    DOI 10.3389/fimmu.2023.1225603
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Complementing antibody profiles: assessing antibody function on antigen microarrays.

    Prechl, József / Szittner, Zoltán / Papp, Krisztián

    Immunology letters

    2012  Volume 143, Issue 1, Page(s) 101–105

    Abstract: Antibody effector functions other than neutralization depend on interactions with soluble and cellular components of the immune system. Antigen recognition is usually oligoclonal, with the different clones of antibodies belonging to different classes, ... ...

    Abstract Antibody effector functions other than neutralization depend on interactions with soluble and cellular components of the immune system. Antigen recognition is usually oligoclonal, with the different clones of antibodies belonging to different classes, subclasses, glycoforms and having different affinities and epitope specificities. Thus, composition of immune complexes determines biological effects mainly via interactions with FcR and complement proteins. Antibodies are capable of triggering any of the three pathways of complement activation and antigen recognition of complex antigens often results in the activation of more than one pathway. These events can be tracked in a multiplex format using antigen microarrays, where complement products bind to elements of the microarray. By controlling cation concentrations and detecting various complement components (C1q, C4, C3) contribution of the different pathways can be identified. Parallel measurement of antibodies and complement proteins provides a novel way of looking at interactions between antigen and antibodies. We propose the use of immune complex signatures, composite depictions of antibody and complement content of immune complexes characterizing healthy and diseased populations. Normalized interquartile ranges of antibody binding (IgM, IgG) and complement deposition (C4, C3) are projected onto radar charts to produce patterns that can distinguish normal and altered immune responses. We propose that comprehensive interaction studies of serum antibodies and complement with arrays of antigens can generate functional antibody profiles and help better understand immunological disease mechanism.
    MeSH term(s) Antigen-Antibody Complex/chemistry ; Antigen-Antibody Complex/immunology ; Antigens/chemistry ; Antigens/immunology ; Complement C1q/chemistry ; Complement C1q/immunology ; Complement C3/chemistry ; Complement C3/immunology ; Complement C4/chemistry ; Complement C4/immunology ; Humans ; Immunoglobulin G/chemistry ; Immunoglobulin G/immunology ; Immunoglobulin M/chemistry ; Immunoglobulin M/immunology ; Models, Molecular ; Protein Array Analysis ; Protein Interaction Domains and Motifs
    Chemical Substances Antigen-Antibody Complex ; Antigens ; Complement C3 ; Complement C4 ; Immunoglobulin G ; Immunoglobulin M ; Complement C1q (80295-33-6)
    Language English
    Publishing date 2012-03-30
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 445150-8
    ISSN 1879-0542 ; 0165-2478
    ISSN (online) 1879-0542
    ISSN 0165-2478
    DOI 10.1016/j.imlet.2012.01.011
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  6. Article ; Online: Life on a microarray: assessing live cell functions in a microarray format.

    Papp, Krisztián / Szittner, Zoltán / Prechl, József

    Cellular and molecular life sciences : CMLS

    2012  Volume 69, Issue 16, Page(s) 2717–2725

    Abstract: Microarray technology outgrew the detection of simple intermolecular interactions, as incubation of slides with living cells opened new vistas. Cell-based array technology permits simultaneous detection of several different cell surface molecules, ... ...

    Abstract Microarray technology outgrew the detection of simple intermolecular interactions, as incubation of slides with living cells opened new vistas. Cell-based array technology permits simultaneous detection of several different cell surface molecules, allowing the complex characterization of cells with an amount of information that is hardly assessed by any other technique. Furthermore, binding of cells to printed antibodies or ligands may induce their activation, and consequently the outcome of these interactions, such as phosphorylation, gene expression, secretion of various products; differentiation, proliferation and apoptosis of the cells are also measurable on arrays. Moreover, since cells can be transfected with printed vectors, over- or under-expression of selected genes is also achievable simultaneously, creating a nice tool for assessing the function of a given gene. The enormously high-throughput cell-based microarray technology enables testing the effect of external stimuli on a scale that was earlier unthinkable. This review summarizes the possible applications of cell-based arrays.
    MeSH term(s) Animals ; Cell Physiological Phenomena ; Drug Evaluation, Preclinical ; High-Throughput Screening Assays ; Humans ; Oligonucleotide Array Sequence Analysis ; Tissue Array Analysis
    Language English
    Publishing date 2012-03-04
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 1358415-7
    ISSN 1420-9071 ; 1420-682X
    ISSN (online) 1420-9071
    ISSN 1420-682X
    DOI 10.1007/s00018-012-0947-z
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  7. Article ; Online: Application of fluorescent monocytes for probing immune complexes on antigen microarrays.

    Szittner, Zoltán / Papp, Krisztián / Sándor, Noémi / Bajtay, Zsuzsa / Prechl, József

    PloS one

    2013  Volume 8, Issue 9, Page(s) e72401

    Abstract: Microarrayed antigens are used for identifying serum antibodies with given specificities and for generating binding profiles. Antibodies bind to these arrayed antigens forming immune complexes and are conventionally identified by secondary labelled ... ...

    Abstract Microarrayed antigens are used for identifying serum antibodies with given specificities and for generating binding profiles. Antibodies bind to these arrayed antigens forming immune complexes and are conventionally identified by secondary labelled antibodies.In the body immune complexes are identified by bone marrow derived phagocytic cells, such as monocytes. In our work we were looking into the possibility of replacing secondary antibodies with monocytoid cells for the generation of antibody profiles. Using the human monocytoid cell line U937, which expresses cell surface receptors for immune complex components, we show that cell adhesion is completely dependent on the interaction of IgG heavy chains and Fcγ receptors, and this recognition is susceptible to differences between heavy chain structures and their glycosylation. We also report data on a possible application of this system in autoimmune diagnostics.Compared to secondary antibodies, fluorescent monocytesas biosensors are superior in reflecting biological functions of microarray-bound antibodies and represent an easy and robust alternative for profiling interactions between serum proteins and antigens.
    MeSH term(s) Antibodies, Immobilized/chemistry ; Autoantigens/chemistry ; Autoantigens/immunology ; Autoantigens/metabolism ; Cell Adhesion ; Cell Line, Tumor ; Complement System Proteins/metabolism ; Fluorescent Dyes/chemistry ; Humans ; Immunoglobulin G/blood ; Immunoglobulin M/blood ; Lupus Erythematosus, Systemic/blood ; Lupus Erythematosus, Systemic/immunology ; Monocytes/immunology ; Protein Array Analysis/methods ; Protein Binding ; Receptors, IgG/metabolism
    Chemical Substances Antibodies, Immobilized ; Autoantigens ; Fluorescent Dyes ; Immunoglobulin G ; Immunoglobulin M ; Receptors, IgG ; Complement System Proteins (9007-36-7)
    Language English
    Publishing date 2013-09-05
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0072401
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  8. Article ; Online: Author Correction: Human DC-SIGN and CD23 do not interact with human IgG.

    Temming, A Robin / Dekkers, Gillian / van de Bovenkamp, Fleur S / Plomp, H Rosina / Bentlage, Arthur E H / Szittner, Zoltán / Derksen, Ninotska I L / Wuhrer, Manfred / Rispens, Theo / Vidarsson, Gestur

    Scientific reports

    2020  Volume 10, Issue 1, Page(s) 12560

    Abstract: An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    Abstract An amendment to this paper has been published and can be accessed via a link at the top of the paper.
    Language English
    Publishing date 2020-07-23
    Publishing country England
    Document type Published Erratum
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-020-68760-2
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  9. Article ; Online: Label-free detection of immune complexes with myeloid cells.

    Szittner, Z / Bentlage, A E H / Rovero, P / Migliorini, P / Lóránd, V / Prechl, J / Vidarsson, G

    Clinical and experimental immunology

    2016  Volume 185, Issue 1, Page(s) 72–80

    Abstract: The aim of this study was to provide proof-of-concept for quantitative and qualitative label-free detection of immune complexes through myeloid cells with imaging surface plasmon resonance. Surface plasmon resonance imaging was first applied to monitor ... ...

    Abstract The aim of this study was to provide proof-of-concept for quantitative and qualitative label-free detection of immune complexes through myeloid cells with imaging surface plasmon resonance. Surface plasmon resonance imaging was first applied to monitor the binding of human sera from healthy and rheumatoid arthritis (RA) patients to immobilized citrullinated RA-specific peptide antigens, histone citrullinated peptide 2 (HCP2) and viral citrullinated peptide 2 (VCP2). Next, the binding of monocytoid cell line U937 to the resulting immune complexes on the sensor surface was monitored. As control, binding of U937 was monitored to immunoglobulin (Ig)G subclasses simultaneously. Cell response results were compared to results of cyclic citrullinated peptide 2 (CCP2) enzyme-linked immunosorbent assay (ELISA), clinical RA diagnosis and antigen-specific antibody distribution of the samples. Human IgG3 triggered the most pronounced response, followed by IgG1 and IgG4, while IgG2 did not result in U937 cell binding. Serum samples obtained from RA patients resulted in a significantly increased cell response to VCP2 compared to healthy controls. The strength of cell response towards VCP2 immune complexes showed significant correlation with levels of antigen-specific IgA, IgG and IgG3. Cellular responses on VCP2 immune complexes showed significant association with both CCP2-based serological positivity and European League Against Rheumatism (EULAR) criteria-based clinical RA diagnosis. Immunoglobulin-triggered binding of monocytoid cells can be monitored using a label-free multiplex technology. Because these binding events are presumably initiated by Fc receptors, the system provides a tool for biological detection of autoantibodies with diagnostic value, here exemplified by anti-citrullinated antibodies. This provides added information to antibody levels, as interaction with Fc-receptor-expressing cells is also affected by post-translational modification of the immunoglobulins.
    MeSH term(s) Antigen-Antibody Complex/analysis ; Antigen-Antibody Complex/metabolism ; Arthritis, Rheumatoid/blood ; Arthritis, Rheumatoid/diagnosis ; Arthritis, Rheumatoid/immunology ; Arthritis, Rheumatoid/pathology ; Autoantibodies/chemistry ; Autoantibodies/metabolism ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immobilized Proteins/blood ; Immobilized Proteins/chemistry ; Immune Sera/chemistry ; Immunoglobulin G/blood ; Peptide Fragments/blood ; Peptide Fragments/chemistry ; Peptides, Cyclic/blood ; Peptides, Cyclic/chemistry ; Protein Binding ; Surface Plasmon Resonance ; U937 Cells ; Viral Proteins/blood ; Viral Proteins/chemistry
    Chemical Substances Antigen-Antibody Complex ; Autoantibodies ; EBNA-2 protein (338-358), citrullinated- ; Immobilized Proteins ; Immune Sera ; Immunoglobulin G ; Peptide Fragments ; Peptides, Cyclic ; Viral Proteins ; cyclic citrullinated peptide
    Language English
    Publishing date 2016
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 218531-3
    ISSN 1365-2249 ; 0009-9104 ; 0964-2536
    ISSN (online) 1365-2249
    ISSN 0009-9104 ; 0964-2536
    DOI 10.1111/cei.12788
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  10. Article ; Online: Immunoassay for quantification of antigen-specific IgG fucosylation.

    Šuštić, Tonći / 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

    EBioMedicine

    2022  Volume 81, Page(s) 104109

    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 ... ...

    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 (R
    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.
    MeSH term(s) COVID-19/diagnosis ; COVID-19/therapy ; Enzyme-Linked Immunosorbent Assay/methods ; Fucose/chemistry ; Fucose/metabolism ; Humans ; Immunization, Passive ; Immunoglobulin Fc Fragments/chemistry ; Immunoglobulin G/chemistry ; Receptors, IgG/chemistry
    Chemical Substances Immunoglobulin Fc Fragments ; Immunoglobulin G ; Receptors, IgG ; Fucose (28RYY2IV3F)
    Language English
    Publishing date 2022-06-22
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2851331-9
    ISSN 2352-3964
    ISSN (online) 2352-3964
    DOI 10.1016/j.ebiom.2022.104109
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