Malaria pp 307-319 | Cite as

Expressing Full-Length Functional PfEMP1 Proteins in the HEK293 Expression System

  • Anand Srivastava
  • Yves Durocher
  • Benoît GamainEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 923)


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.

Key words

Plasmodium falciparum Malaria PfEMP1 var var2CSA HEK293 expression system 



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).


  1. 1.
    Florens L et al (2002) A proteomic view of the Plasmodium falciparum life cycle. Nature 419:520–526PubMedCrossRefGoogle Scholar
  2. 2.
    Gardner MJ et al (2002) Genome sequence of the human malaria parasite Plasmodium falciparum. Nature 419:498–511PubMedCrossRefGoogle Scholar
  3. 3.
    Durocher Y, Butler M (2009) Expression systems for therapeutic glycoprotein production. Curr Opin Biotechnol 20:700–707PubMedCrossRefGoogle Scholar
  4. 4.
    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
  5. 5.
    Wurm F, Bernard A (1999) Large-scale transient expression in mammalian cells for recombinant protein production. Curr Opin Biotechnol 10:156–159PubMedCrossRefGoogle Scholar
  6. 6.
    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
  7. 7.
    Geisse S (2009) Reflections on more than 10 years of TGE approaches. Protein Expr Purif 64:99–107PubMedCrossRefGoogle Scholar
  8. 8.
    Pham PL et al (2006) Large-scale transfection of mammalian cells for the fast production of recombinant protein. Mol Biotechnol 34:225–237PubMedCrossRefGoogle Scholar
  9. 9.
    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
  10. 10.
    Sambrook J et al (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  11. 11.
    Zhang J et al (2009) Transient expression and purification of chimeric heavy chain antibodies. Protein Expr Purif 65:77–82PubMedCrossRefGoogle Scholar
  12. 12.
    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

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Anand Srivastava
    • 1
  • Yves Durocher
    • 2
  • Benoît Gamain
    • 1
    Email author
  1. 1.Institut National de Transfusion SanguineUniversité Paris DiderotParisFrance
  2. 2.Biotechnology Research Institute, National Research Council CanadaMontrealCanada

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