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Published on: January 3, 2012
Transfected HEK293 cells expressing functional recombinant intercellular adhesion molecule 1 (ICAM-1)--a receptor
Anja Bengtsson1, Louise Joergensen, Zachary R Barbati
1Centre for Medical Parasitology at the Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen and at Department of Infectious Diseases, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
Abstract:
Intercellular adhesion molecule 1 (ICAM-1) is a membrane-bound glycoprotein expressed on endothelial cells and cells of the immune system. Human ICAM-1 mediates adhesion and migration of leucocytes, and is implicated in inflammatory pathologies, autoimmune diseases and in many cancer processes. Additionally, ICAM-1 acts as receptor for pathogens like human rhinovirus and Plasmodium falciparum malaria parasites. A group of related P. falciparum erythrocyte membrane protein 1 (PfEMP1) domains, the DBLβ, mediates ICAM-1 binding of P. falciparum-infected erythrocytes. This ICAM‑1-binding phenotype has been suggested to be involved in the development of cerebral malaria. However, more studies identifying cross-reactive antibody and ICAM-1-binding epitopes and the establishment of a clinical link between DBLβ expression and e.g. cerebral malaria are needed before the DBLβ domains can be put forward as vaccine candidates and go into clinical trials. Such studies require availability of functional recombinant ICAM-1 in large quantities. In this study, we compared recombinant ICAM-1 expressed in HEK293 and COS-7 cells with mouse myeloma NS0 ICAM-1 purchased from a commercial vendor in terms of protein purity, yield, fold, ability to bind DBLβ, and relative cost. We present a HEK293 cell-based, high-yield expression and purification scheme for producing inexpensive, functional ICAM‑1. ICAM-1 expressed in HEK293 is applicable to malaria research and can also be useful in other research fields.
Insights
We developed a cost-effective method to produce functional recombinant Intercellular Adhesion Molecule 1 (ICAM-1) in HEK293 cells. This ICAM-1 is crucial for malaria research and other fields requiring this important protein.
Area of Science:
- Immunology
- Molecular Biology
- Parasitology
Background:
- Intercellular Adhesion Molecule 1 (ICAM-1) is a key protein in immune responses and pathogen binding.
- ICAM-1 binding by Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) domains (DBLβ) is linked to severe malaria.
- Functional recombinant ICAM-1 is needed for vaccine development and further research.
Purpose of the Study:
- To compare different methods of producing recombinant ICAM-1.
- To establish a high-yield, cost-effective expression and purification scheme for functional ICAM-1.
- To provide a valuable tool for malaria research and other scientific fields.
Main Methods:
- Recombinant ICAM-1 was expressed in HEK293 and COS-7 cells.
- Commercial ICAM-1 from NS0 cells was used as a benchmark.
- Protein purity, yield, fold, DBLβ binding ability, and cost were evaluated.
Main Results:
- A HEK293 cell-based expression and purification scheme was successfully developed.
- This method yields inexpensive and functional recombinant ICAM-1.
- The produced ICAM-1 demonstrated ability to bind DBLβ domains.
Conclusions:
- The developed HEK293 cell-based system provides a scalable and economical source of functional ICAM-1.
- This recombinant ICAM-1 is suitable for malaria research, particularly for studying ICAM-1 binding phenotypes.
- The method offers a valuable resource for broader research applications beyond malaria.
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