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The Plasmodium falciparum circumsporozoite protein produced in Lactococcus lactis is pure and stable
- Source :
- Singh, S K, Plieskatt, J, Chourasia, B K, Singh, V, Bolscher, J M, Dechering, K J, Adu, B, López-Méndez, B, Kaviraj, S, Locke, E, King, C R & Theisen, M 2020, ' The Plasmodium falciparum circumsporozoite protein produced in Lactococcus lactis is pure and stable ', Journal of Biological Chemistry, vol. 295, no. 2, pp. 403-414 . https://doi.org/10.1074/jbc.RA119.011268
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- The Plasmodium falciparum circumsporozoite protein (PfCSP) is a sporozoite surface protein whose role in sporozoite motility and cell invasion has made it the leading candidate for a pre-erythrocytic malaria vaccine. However, production of high yields of soluble recombinant PfCSP, including its extensive NANP and NVDP repeats, has proven problematic. Here, we report on the development and characterization of a secreted, soluble, and stable full-length PfCSP (containing 4 NVDP and 38 NANP repeats) produced in the Lactococcus lactis expression system. The recombinant full-length PfCSP, denoted PfCSP4/38, was produced initially with a histidine tag and purified by a simple two-step procedure. Importantly, the recombinant PfCSP4/38 retained a conformational epitope for antibodies as confirmed by both in vivo and in vitro characterizations. We characterized this complex protein by HPLC, light scattering, MS analysis, differential scanning fluorimetry, CD, SDS-PAGE, and immunoblotting with conformation-dependent and -independent mAbs, which confirmed it to be both pure and soluble. Moreover, we found that the recombinant protein is stable at both frozen and elevated-temperature storage conditions. When we used L. lactis-derived PfCSP4/38 to immunize mice, it elicited high levels of functional antibodies that had the capacity to modify sporozoite motility in vitro We concluded that the reported yield, purity, results of biophysical analyses, and stability of PfCSP4/38 warrant further consideration of using the L. lactis system for the production of circumsporozoite proteins for preclinical and clinical applications in malaria vaccine development.
- Subjects :
- 0301 basic medicine
030102 biochemistry & molecular biology
biology
Chemistry
Malaria vaccine
Immunogenicity
Lactococcus lactis
Plasmodium falciparum
Cell Biology
biology.organism_classification
Biochemistry
In vitro
law.invention
Circumsporozoite protein
03 medical and health sciences
030104 developmental biology
law
Recombinant DNA
Molecular Biology
Conformational epitope
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 295
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi.dedup.....ed5e916b53edf5b6e352338d21bb7759