High-Throughput BioPlex Assay for the Study of Functionally Active Plasmodium Falciparum Antigens That Are Expressed on the Surface of Infected Erythrocytes.

Identification of P. falciparum infected erythrocyte surface ligands (such as PfEMP1) matched with the host receptors they interact with, as well as identification of PfEMP1 domains that are targets of protective immunity, are important for understanding of the pathophysiology of severe malaria (SM) and for design of novel vaccine candidates. In addition, identification of small-molecule drugs that can prevent or reverse receptor-ligand domain interactions could provide new tools for adjunctive therapy in SM. This protocol describes how to prepare functionally intact PfEMP1 proteins in mammalian cells (COS-7) and immobilize them on the surface of BioPlex beads. Furthermore, the protocol described how to identify PfEMP1 constructs that bind to specific host receptors or to immunoglobulins (IgG, IgM, etc.), and how to measure inhibition of the receptor binding to PfEMP1 constructs by small-molecule compounds or serum/plasma.