Levels of VAR2CSA-specific IgG were assessed by ELISA as previously described68 (link). In brief, 96-well flat-bottom microtiter plates (Nunc MaxiSorp, Thermo Fisher Scientific) were coated overnight at 4 °C with full-length VAR2CSA (100 ng/well in PBS). Monoclonal antibody (0.08–10 μg/mL) or plasma samples (1:400) were added in duplicate, followed by washing and horseradish peroxidase-conjugated rabbit anti-human IgG (1:3,000; Dako). Bound antibodies were detected by adding TMB PLUS2 (Eco-Tek), and the reaction stopped by the addition of 0.2 M H2SO4. The optical density (OD) was read at 450 nm and the specific antibody levels were calculated in arbitrary units (arb. units), using the equation 100 × [(ODSAMPLE − ODBLANK)/(ODPOS.CTRL − ODBLANK)].
Larsen M.D., Lopez-Perez M., Dickson E.K., Ampomah P., Tuikue Ndam N., Nouta J., Koeleman C.A., Ederveen A.L., Mordmüller B., Salanti A., Nielsen M.A., Massougbodji A., van der Schoot C.E., Ofori M.F., Wuhrer M., Hviid L, & Vidarsson G. (2021). Afucosylated Plasmodium falciparum-specific IgG is induced by infection but not by subunit vaccination. Nature Communications, 12, 5838.
Other organizations :
Sanquin, University of Copenhagen, Noguchi Memorial Institute for Medical Research, University of Ghana, University of Cape Coast, Mère et Enfant en Milieu Tropical, Université Paris Cité, Institut de Recherche pour le Développement, Netherlands Metabolomics Centre, Leiden University Medical Center, Radboud University Nijmegen, Radboud University Medical Center, Université d'Abomey-Calavi, University of Amsterdam, Amsterdam University Medical Centers
Levels of VAR2CSA-specific IgG (measured in arbitrary units)
control variables
Coating of 96-well plates with full-length VAR2CSA (100 ng/well in PBS)
Washing of plates
Addition of horseradish peroxidase-conjugated rabbit anti-human IgG (1:3,000)
Detection of bound antibodies using TMB PLUS2 and 0.2 M H2SO4
controls
Positive control: Not explicitly mentioned
Negative control: Blank (OD_BLANK)
Annotations
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