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Updated: Sep 3, 2025
High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension
Published on: December 28, 2015
Expression of Large Full-Length PfEMP1 Proteins in HEK293 Cells
Jonathan Paul Renn1, Justin Yai Alamou Doritchamou1, Patrick Emmet Duffy2
1Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
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.
Insights
Researchers developed an improved method to produce more full-length VAR2CSA, a key protein for malaria vaccines. This advancement facilitates greater quantities of the protein for structural and functional studies, aiding vaccine development against pregnancy malaria.
Area of Science:
- Parasitology
- Vaccinology
- Protein Expression
Background:
- Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is crucial for parasite virulence and severe malaria.
- VAR2CSA, a specific PfEMP1, is essential for pregnancy malaria and a vaccine target.
- Recombinant full-length VAR2CSA expression is challenging due to its size, complexity, and high cysteine content.
Purpose of the Study:
- To enhance the yield of full-length VAR2CSA for improved protein characterization.
- To develop a modified protocol for higher quantity production of VAR2CSA.
Main Methods:
- Utilized advanced mammalian expression systems.
- Modified existing protocols to overcome expression challenges.
- Focused on optimizing protein yields for structural and functional studies.
Main Results:
- Achieved enhanced yields of full-length VAR2CSA beyond current systems.
- Generated higher quantities of recombinant VAR2CSA protein.
- Enabled more extensive protein structural and functional characterization.
Conclusions:
- The modified protocol significantly improves full-length VAR2CSA production.
- Increased protein availability supports further research into VAR2CSA's structure and function.
- This advancement is critical for developing effective pregnancy malaria vaccines.
