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Expression of Large Full-Length PfEMP1 Proteins in HEK293 Cells

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Malaria Immunology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2470))

Abstract

Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is a family of proteins expressed on the surface of red blood cells infected by Plasmodium falciparum. PfEMP1 proteins play a vital role in parasite virulence, and thus are important vaccine candidates to prevent severe disease. VAR2CSA is one specific PfEMP1 essential for pregnancy malaria pathogenesis, and the primary target in pregnancy malaria vaccine development. However, similar to other PfEMP1 proteins, expression of recombinant full-length VAR2CSA is difficult due to its large size, multidomain architecture and high cysteine content. To date, there has been success using higher ordered expression systems (such as mammalian and insect cells) to generate folded and active VAR2CSA. However, recent improvements with mammalian expression systems including cell lines and promoters have pushed the boundaries of yields. Here, we describe a modified protocol beyond current systems that enhances yields of full-length VAR2CSA and can generate higher quantities of material for protein structural and functional characterization.

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Acknowledgments

This work was supported by the Intramural Research Program of the National Institute of Allergy and Infectious Diseases, National Institutes of Health. We thank J. Patrick Gorres for assistance in editing the manuscript.

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Authors and Affiliations

  1. Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA

    Jonathan Paul Renn, Justin Yai Alamou Doritchamou & Patrick Emmet Duffy

Authors
  1. Jonathan Paul Renn
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  2. Justin Yai Alamou Doritchamou
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  3. Patrick Emmet Duffy
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Correspondence to Patrick Emmet Duffy .

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Editors and Affiliations

  1. Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark

    Anja Tatiana Ramstedt Jensen

  2. Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark

    Lars Hviid

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Renn, J.P., Doritchamou, J.Y.A., Duffy, P.E. (2022). Expression of Large Full-Length PfEMP1 Proteins in HEK293 Cells. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_21

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  • DOI: https://doi.org/10.1007/978-1-0716-2189-9_21

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2188-2

  • Online ISBN: 978-1-0716-2189-9

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