Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay
The issue of antibody cross-reactivity is of central importance in immunology, and not least in protective immunity to Plasmodium falciparum malaria, where key antigens show substantial allelic variation (polymorphism). However, serological analysis often does not allow the distinction between true...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2022-07-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/79245 |
| _version_ | 1797998183946649600 |
|---|---|
| author | Henriette Hoffmann-Veltung Nsoh Godwin Anabire Michael Fokuo Ofori Peter Janhmatz Niklas Ahlborg Lars Hviid Maria del Pilar Quintana |
| author_facet | Henriette Hoffmann-Veltung Nsoh Godwin Anabire Michael Fokuo Ofori Peter Janhmatz Niklas Ahlborg Lars Hviid Maria del Pilar Quintana |
| author_sort | Henriette Hoffmann-Veltung |
| collection | DOAJ |
| description | The issue of antibody cross-reactivity is of central importance in immunology, and not least in protective immunity to Plasmodium falciparum malaria, where key antigens show substantial allelic variation (polymorphism). However, serological analysis often does not allow the distinction between true cross-reactivity (one antibody recognizing multiple antigen variants) and apparent cross-reactivity (presence of multiple variant-specific antibodies), as it requires analysis at the single B-cell/monoclonal antibody level. ELISpot is an assay that enables that, and a recently developed multiplexed variant of ELISpot (FluoroSpot) facilitates simultaneous assessment of B-cell/antibody reactivity to several different antigens. In this study, we present a further enhancement of this assay that makes direct analysis of monoclonal antibody-level cross-reactivity with allelic variants feasible. Using VAR2CSA-type PfEMP1—a notoriously polymorphic antigen involved in the pathogenesis of placental malaria—as a model, we demonstrate the robustness of the assay and its applicability to analysis of true cross-reactivity of monoclonal VAR2CSA-specific antibodies in naturally exposed individuals. The assay is adaptable to the analysis of other polymorphic antigens, rendering it a powerful tool in studies of immunity to malaria and many other diseases. |
| first_indexed | 2024-04-11T10:44:39Z |
| format | Article |
| id | doaj.art-9e7e19c7292f4b0192ea86e0adeac26f |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T10:44:39Z |
| publishDate | 2022-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-9e7e19c7292f4b0192ea86e0adeac26f2022-12-22T04:29:05ZengeLife Sciences Publications LtdeLife2050-084X2022-07-011110.7554/eLife.79245Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assayHenriette Hoffmann-Veltung0Nsoh Godwin Anabire1Michael Fokuo Ofori2Peter Janhmatz3Niklas Ahlborg4Lars Hviid5https://orcid.org/0000-0002-1698-4927Maria del Pilar Quintana6https://orcid.org/0000-0001-6190-3324Centre for Medical Parasitology, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkCentre for Medical Parasitology, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology, University of Ghana, Accra, Ghana; Department of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, GhanaDepartment of Immunology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, GhanaMabtech AB, Nacka Strand, SwedenMabtech AB, Nacka Strand, Sweden; Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm, SwedenCentre for Medical Parasitology, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Department of Infectious Diseases, Rigshospitalet, Copenhagen, DenmarkCentre for Medical Parasitology, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkThe issue of antibody cross-reactivity is of central importance in immunology, and not least in protective immunity to Plasmodium falciparum malaria, where key antigens show substantial allelic variation (polymorphism). However, serological analysis often does not allow the distinction between true cross-reactivity (one antibody recognizing multiple antigen variants) and apparent cross-reactivity (presence of multiple variant-specific antibodies), as it requires analysis at the single B-cell/monoclonal antibody level. ELISpot is an assay that enables that, and a recently developed multiplexed variant of ELISpot (FluoroSpot) facilitates simultaneous assessment of B-cell/antibody reactivity to several different antigens. In this study, we present a further enhancement of this assay that makes direct analysis of monoclonal antibody-level cross-reactivity with allelic variants feasible. Using VAR2CSA-type PfEMP1—a notoriously polymorphic antigen involved in the pathogenesis of placental malaria—as a model, we demonstrate the robustness of the assay and its applicability to analysis of true cross-reactivity of monoclonal VAR2CSA-specific antibodies in naturally exposed individuals. The assay is adaptable to the analysis of other polymorphic antigens, rendering it a powerful tool in studies of immunity to malaria and many other diseases.https://elifesciences.org/articles/79245antibodycross-reactivityfluorospot |
| spellingShingle | Henriette Hoffmann-Veltung Nsoh Godwin Anabire Michael Fokuo Ofori Peter Janhmatz Niklas Ahlborg Lars Hviid Maria del Pilar Quintana Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay eLife antibody cross-reactivity fluorospot |
| title | Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay |
| title_full | Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay |
| title_fullStr | Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay |
| title_full_unstemmed | Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay |
| title_short | Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay |
| title_sort | analysis of allelic cross reactivity of monoclonal igg antibodies by a multiplexed reverse fluorospot assay |
| topic | antibody cross-reactivity fluorospot |
| url | https://elifesciences.org/articles/79245 |
| work_keys_str_mv | AT henriettehoffmannveltung analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT nsohgodwinanabire analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT michaelfokuoofori analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT peterjanhmatz analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT niklasahlborg analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT larshviid analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay AT mariadelpilarquintana analysisofalleliccrossreactivityofmonoclonaliggantibodiesbyamultiplexedreversefluorospotassay |