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Media type:
E-Article
Title:
Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay
Contributor:
Hoffmann-Veltung, Henriette;
Anabire, Nsoh Godwin;
Ofori, Michael Fokuo;
Janhmatz, Peter;
Ahlborg, Niklas;
Hviid, Lars;
Quintana, Maria del Pilar
imprint:
eLife Sciences Publications, Ltd, 2022
Published in:eLife
Language:
English
DOI:
10.7554/elife.79245
ISSN:
2050-084X
Origination:
Footnote:
Description:
<jats:p>The issue of antibody cross-reactivity is of central importance in immunology, and not least in protective immunity to <jats:italic>Plasmodium falciparum</jats:italic> 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.</jats:p>