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Published on: July 17, 2014
Large-scale purification and characterization of malaria vaccine candidate antigen Pvs25H for use in clinical trials
Aaron P Miles1, Yanling Zhang, Allan Saul
1Malaria Vaccine Development Unit, National Institute of Allergy and Infectious Diseases, Rockville, Maryland 20852, USA.
Abstract:
The budding yeast Saccharomyces cerevisiae has been used to express the recombinant protein Pvs25H, currently the only candidate transmission-blocking vaccine against Plasmodium vivax malaria. This molecule contains four epidermal growth factor-like domains and is expressed as at least two stable monomeric forms with different physicochemical properties. Pvs25H-A is apparently homogeneous and seems to have a correct disulfide bond structure. By contrast, Pvs25H-B is produced as a heterogeneous population of molecules, some of which are associated with an as yet unidentified chromophore, and it contains both internal and N-terminal cleavages. We report here a procedure for successfully separating these two forms with a process suitable for clinical production of this antigen.
Insights
Researchers developed a method to separate two forms of Pvs25H, a malaria vaccine candidate. This process is suitable for the clinical production of the Plasmodium vivax antigen.
Area of Science:
- Biotechnology
- Vaccine Development
- Parasitology
Background:
- The budding yeast Saccharomyces cerevisiae is utilized for recombinant protein expression.
- Pvs25H is a candidate transmission-blocking vaccine against Plasmodium vivax malaria.
- Pvs25H exhibits distinct monomeric forms with differing properties.
Purpose of the Study:
- To develop a method for separating the two stable monomeric forms of Pvs25H.
- To ensure the clinical production of the malaria antigen.
Main Methods:
- Expression of recombinant Pvs25H in Saccharomyces cerevisiae.
- Characterization of Pvs25H monomeric forms (Pvs25H-A and Pvs25H-B).
- Development of a separation procedure for clinical-scale production.
Main Results:
- Pvs25H-A is a homogeneous form with correct disulfide bonds.
- Pvs25H-B is heterogeneous, associated with a chromophore, and contains cleavages.
- A successful procedure for separating Pvs25H-A and Pvs25H-B was established.
Conclusions:
- The developed procedure enables the separation of Pvs25H forms.
- This method is suitable for the clinical production of the Pvs25H antigen for malaria vaccine development.
