Forskning
Udskriv Udskriv
Switch language
Region Hovedstaden - en del af Københavns Universitetshospital
Udgivet

Analysis of allelic cross-reactivity of monoclonal IgG antibodies by a multiplexed reverse FluoroSpot assay

Publikation: Bidrag til tidsskriftTidsskriftartikelpeer review

DOI

  • Henriette Hoffmann-Veltung
  • Nsoh Godwin Anabire
  • Michael Fokuo Ofori
  • Peter Janhmatz
  • Niklas Ahlborg
  • Lars Hviid
  • Maria Del Pilar Quintana
Vis graf over relationer

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.

OriginalsprogEngelsk
Artikelnummere79245
TidsskrifteLife
Vol/bind11
ISSN2050-084X
DOI
StatusUdgivet - 15 jul. 2022

Bibliografisk note

© 2022, Hoffmann-Veltung et al.

ID: 79467133