Due to the A/T-richness of the genome of Plasmodium falciparum, expressing P. falciparum proteins in heterologous expression systems is challenging. In addition, many P. falciparum proteins have high cysteine content and high molecular weight, which further complicates expression of these proteins in heterologous systems. The high molecular weight Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1) adhesins expressed on the surface of the infected erythrocytes are among the most difficult proteins to express. Cost reduction in synthetic gene synthesis, as well as improved eukaryotic expression systems, now makes it possible to express such proteins. In this chapter, we describe the construction, production, purification, and functional assessment of the full-length extracellular region of the var2CSA PfEMP1 protein involved in pregnancy-associated malaria (PAM), using a human embryonic kidney (HEK) expression system.
Plasmodium falciparumMalaria PfEMP1 varvar2CSA HEK293 expression system
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The research leading to these results has received funding from the European Community’s Seventh Framework Programme Grant ([FP7/2007-2013]) under Grant agreement 201222. A.S. is supported by a grant from the Fondation pour la Recherche Médicale (FRM) (SPF20101220957).
Durocher Y, Butler M (2009) Expression systems for therapeutic glycoprotein production. Curr Opin Biotechnol 20:700–707PubMedCrossRefGoogle Scholar
Graham FL et al (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74PubMedCrossRefGoogle Scholar
Wurm F, Bernard A (1999) Large-scale transient expression in mammalian cells for recombinant protein production. Curr Opin Biotechnol 10:156–159PubMedCrossRefGoogle Scholar
Baldi L et al (2007) Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectives. Biotechnol Lett 29:677–684PubMedCrossRefGoogle Scholar
Pham PL et al (2006) Large-scale transfection of mammalian cells for the fast production of recombinant protein. Mol Biotechnol 34:225–237PubMedCrossRefGoogle Scholar
Durocher Y et al (2002) High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res 30:e9PubMedCrossRefGoogle Scholar
Sambrook J et al (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
Zhang J et al (2009) Transient expression and purification of chimeric heavy chain antibodies. Protein Expr Purif 65:77–82PubMedCrossRefGoogle Scholar
Srivastava A et al (2010) Full-length extracellular region of the var2CSA variant of PfEMP1 is required for specific, high-affinity binding to CSA. Proc Natl Acad Sci USA 107:4884–4889PubMedCrossRefGoogle Scholar