• Authors: Kucharska I. et al.
  • Year: 2022
  • Journal: Elife 11 e72908
  • Applications: in vitro / DNA / FectoPRO
  • Cell type: HEK-293F


pcDNA3.4-Fab KC and Fab HC plasmids were co-transfected into HEK 293F cells for transient expression using FectoPRO DNA transfection reagent (Polyplus). Cells were cultured in Gibco FreeStyle 293 Expression Medium for 6–7 days and subsequently purified via a combination of KappaSelect affinity chromatography (GE Healthcare), cation exchange chromatography (MonoS, GE Healthcare), and size-exclusion chromatography (Superdex 200 Increase 10/300 GL, GE Healthcare). The resulting pcDNA3.4-PvCSPvk210-His6x and pcDNA3.4-PvCSPvk247-His6x plasmids were transiently transfected in HEK 293F cells using FectoPRO DNA transfection reagent (Polyplus), cultured in Gibco FreeStyle 293 Expression Medium, and purified by HisTrap FF affinity chromatography (GE Healthcare) and size-exclusion chromatography (Superdex 200 Increase 10/300 GL, GE Healthcare).


Malaria is a global health burden, with Plasmodium falciparum (Pf) and Plasmodium vivax (Pv) responsible for the majority of infections worldwide. Circumsporozoite protein (CSP) is the most abundant protein on the surface of Plasmodium sporozoites, and antibodies targeting the central repeat region of CSP can prevent parasite infection. Although much has been uncovered about the molecular basis of antibody recognition of the PfCSP repeats, data remains scarce for PvCSP. Here, we performed molecular dynamics simulations for peptides comprising the PvCSP repeats from strains VK210 and VK247 to reveal how the PvCSP central repeats are highly disordered, with minor propensities to adopt turn conformations. Next, we solved eight crystal structures to unveil the interactions of two inhibitory monoclonal antibodies (mAbs), 2F2 and 2E10.E9, with PvCSP repeats. Both antibodies can accommodate subtle sequence variances in the repeat motifs and recognize largely coiled peptide conformations that also contain isolated turns. Our structural studies uncover various degrees of Fab-Fab homotypic interactions upon recognition of the PvCSP central repeats by these two inhibitory mAbs, similar to potent mAbs against PfCSP. These findings augment our understanding of host-Plasmodium interactions and contribute molecular details of Pv inhibition by mAbs to unlock structure-based engineering of PvCSP-based vaccines.