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The Plasmodium falciparum circumsporozoite protein produced in Lactococcus lactis is pure and stable
| Content Provider | Scilit |
|---|---|
| Author | Singh, Susheel K. Chourasia, Bishwanath Kumar Singh, Vandana Bolscher, Judith M. Dechering, Koen J. Adu, Bright López-Méndez, Blanca Kaviraj, Swarnendu Locke, Emily King, C. Richter Theisen, Michael |
| Copyright Year | 2020 |
| Description | Journal: Journal of Biological Chemistry 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. |
| Related Links | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6956534/pdf http://www.jbc.org/content/295/2/403.full.pdf |
| Ending Page | 414 |
| Page Count | 12 |
| Starting Page | 403 |
| ISSN | 00219258 |
| e-ISSN | 1083351X |
| DOI | 10.1074/jbc.ra119.011268 |
| Journal | Journal of Biological Chemistry |
| Issue Number | 2 |
| Volume Number | 295 |
| Language | English |
| Publisher | Elsevier BV |
| Publisher Date | 2020-01-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Biological Chemistry Tropical Medicine Lactococcus Lactis Plasmodium Falciparum Circumsporozoite Protein Protein Expression Vaccine Development |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
