Plasmodium falciparum parasites express variable surface antigens on the infected erythrocyte surface allowing adhesion to human host receptors on the blood and endothelial cells, which can result in immune evasion. One of the most studied and key antigens in adhesion is the highly polymorphic PfEMP1. However, despite the vast variation in the PfEMP1 antigens, they are the main targets of naturally acquired immunity and are therefore promising candidates for malaria vaccine development. Generating PfEMP1-specific human monoclonal antibodies from naturally immune individuals will help to determine the best targets of protection from clinical disease. Immortalization of human B cells is one of the oldest and most efficient techniques to generate human monoclonal antibodies. Nevertheless, most protocols require flow cytometry-based cell sorting, which can be a limiting factor for many laboratories. This chapter describes an efficient protocol for the generation of PfEMP1-specific human monoclonal antibodies from malaria immune individuals that can be performed without the use of advanced cell-sorting techniques.
|Tidsskrift||Methods in Molecular Biology|
|Status||Udgivet - 2022|