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Optimization of Expression and Purification of Schistosoma mansoni Antigens in Fusion with Rhizavidin
- Source :
- Molecular Biotechnology
- Publication Year :
- 2021
- Publisher :
- Springer Science and Business Media LLC, 2021.
-
Abstract
- Schistosomiasis causes significant morbidity and mortality. Vaccine efforts to date indicate the need to increase the immunogenicity of Schistosoma antigens. The multiple antigen-presenting system, whereby proteins are genetically fused to rhizavidin and affinity linked to biotinylated templates, enables the generation of robust immune responses. The objective of this work was to express and purify the S. mansoni antigens, SmTSP-2 and SmCD59.2, in fusion with rhizavidin. The fusion with rhizavidin greatly decreased the expression level of rSmTSP-2, but not rSmCD59.2, and both were expressed in the insoluble fraction, requiring optimization of culture conditions. Evaluation of different E. coli strains and media showed that BL21-DE3 cultured in Terrific Broth provided the highest expression levels of both proteins. Investigation of a range of time and temperature of induction showed that E. coli strains expressing rRzv:SmTSP-2 and rRzv:SmCD59.2 showed the highest protein production at 23 °C for 15 h. Recombinant proteins were purified by a single step of affinity chromatography allowing isolation of these proteins in high concentration and purity. The optimization process increased final soluble protein yield of rRzv:SmTSP-2 by fourfold and rRzv:SmCD59.2 by tenfold, providing ~ 20 mg/L of each protein. Optimized fusion protein production will allow antigen use in biotin–rhizavidin affinity platforms. Supplementary Information The online version contains supplementary material available at 10.1007/s12033-021-00355-2.
- Subjects :
- 0106 biological sciences
Recombinant Fusion Proteins
Bioengineering
01 natural sciences
Applied Microbiology and Biotechnology
Biochemistry
Chromatography, Affinity
law.invention
03 medical and health sciences
Bacterial Proteins
Antigen
Affinity chromatography
law
010608 biotechnology
Rhizavidin–biotin system
Protein purification
Animals
Molecular Biology
030304 developmental biology
Original Paper
0303 health sciences
biology
Chemistry
Immunogenicity
Schistosoma mansoni
biology.organism_classification
Fusion protein
Schistosomiasis mansoni
Fusion proteins
Antigens, Helminth
Biotinylation
Recombinant DNA
Protein expression
Schistosoma proteins
Biotechnology
Subjects
Details
- ISSN :
- 15590305 and 10736085
- Volume :
- 63
- Database :
- OpenAIRE
- Journal :
- Molecular Biotechnology
- Accession number :
- edsair.doi.dedup.....f6e31fb6c4f01b9c7b79ebc8c2632e6f