Pe anti human cd54

To measure the ability of an individual’s plasma to inhibit the interaction between the recombinant PfEMP1 DBLβ domain (PF3D7_1150400/PF11_0521) and ICAM1, we used competition ELISA. First, Immunolon plates (Thermo Scientific) were coated with 5µg/mL of PF3D7_1150400 recombinant protein and incubated overnight at 4°C. The plates were then washed and blocked with PBST and 1% skimmed milk for 1h at room temperature. After washing, ICAM1–Fc (5µg/mL) and plasma (1:10) were added simultaneously and incubated for 1h at room temperature (42 (link)). The plates were then washed four times with PBST. Bound ICAM1 was detected by mouse anti-human ICAM-1 (PE anti-human CD54, cat.no 353106, BioLegend, 1:500) followed by goat anti-mouse IgG-HRP, (Dako, 1:5000). Plasma samples from malaria naïve individuals from the UK were used as negative controls.