Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes
Domain III (DIII) of the tick-borne encephalitis virus (TBEV) protein E contains epitopes, which induce antibodies capable of neutralizing the virus. To enhance the immunogenicity of this protein, which has a low molecular weight, the aim of the present work was to express, isolate, and characterize...
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MDPI AG
2018-08-01
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| Series: | Biomolecules |
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| Online Access: | http://www.mdpi.com/2218-273X/8/3/82 |
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| author | Vasily Golotin Nina Sanina Ludmila Davydova Natalia Chopenko Andrey Mazeika Manuel Roig Valery Shnyrov Vladimir N. Uversky Eduard Kostetsky |
| author_facet | Vasily Golotin Nina Sanina Ludmila Davydova Natalia Chopenko Andrey Mazeika Manuel Roig Valery Shnyrov Vladimir N. Uversky Eduard Kostetsky |
| author_sort | Vasily Golotin |
| collection | DOAJ |
| description | Domain III (DIII) of the tick-borne encephalitis virus (TBEV) protein E contains epitopes, which induce antibodies capable of neutralizing the virus. To enhance the immunogenicity of this protein, which has a low molecular weight, the aim of the present work was to express, isolate, and characterize a chimeric protein based on the fusion of the bacterial chaperone HSP70 of Yersinia pseudotuberculosis and EIII (DIII + stem) as a prospective antigen for an adjuvanted delivery system, the tubular immunostimulating complex (TI-complex). The chimeric construction was obtained using pET-40b(+) vector by ligating the respective genes. The resulting plasmid was transformed into DE3 cells for the heterologous expression of the chimeric protein, which was purified by immobilized metal affinity chromatography (IMAC). ELISA, differential scanning calorimetry, intrinsic fluorescence, and computational analysis were applied for the characterization of the immunogenicity and conformation of the chimeric protein. Mice immunization showed that the chimeric protein induced twice the number of anti-EIII antibodies in comparison with EIII alone. In turn, the incorporation of the HSP70/EIII chimeric protein in the TI-complex resulted in a twofold increase in its immunogenicity. The formation of this vaccine construction was accompanied by significant conformational changes in the chimeric protein. Using HSP70 in the content of the chimeric protein represents an efficient means for presenting the main antigenic domain of the TBEV envelope protein to the immune system, whereas the incorporation of this chimeric protein into the TI-complex further contributes to the development of a stronger immune response against the TBEV infection. |
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| language | English |
| last_indexed | 2024-12-11T04:24:22Z |
| publishDate | 2018-08-01 |
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| series | Biomolecules |
| spelling | doaj.art-58bec1758f3d43b0ab1f0804f9010cad2022-12-22T01:21:00ZengMDPI AGBiomolecules2218-273X2018-08-01838210.3390/biom8030082biom8030082Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-ComplexesVasily Golotin0Nina Sanina1Ludmila Davydova2Natalia Chopenko3Andrey Mazeika4Manuel Roig5Valery Shnyrov6Vladimir N. Uversky7Eduard Kostetsky8Department of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDepartment of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDepartment of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDepartment of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDepartment of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDepartamento de Química Física, Facultad de Ciencias Químicas, Universidad de Salamanca, Plaza de los Caìdos s/n, 37008 Salamanca, SpainDepartamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, SpainDepartment of Molecular Medicine and USF Health Byrd Alzheimer’s Research Institute, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd. MDC07, Tampa, FL 33612, USADepartment of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, RussiaDomain III (DIII) of the tick-borne encephalitis virus (TBEV) protein E contains epitopes, which induce antibodies capable of neutralizing the virus. To enhance the immunogenicity of this protein, which has a low molecular weight, the aim of the present work was to express, isolate, and characterize a chimeric protein based on the fusion of the bacterial chaperone HSP70 of Yersinia pseudotuberculosis and EIII (DIII + stem) as a prospective antigen for an adjuvanted delivery system, the tubular immunostimulating complex (TI-complex). The chimeric construction was obtained using pET-40b(+) vector by ligating the respective genes. The resulting plasmid was transformed into DE3 cells for the heterologous expression of the chimeric protein, which was purified by immobilized metal affinity chromatography (IMAC). ELISA, differential scanning calorimetry, intrinsic fluorescence, and computational analysis were applied for the characterization of the immunogenicity and conformation of the chimeric protein. Mice immunization showed that the chimeric protein induced twice the number of anti-EIII antibodies in comparison with EIII alone. In turn, the incorporation of the HSP70/EIII chimeric protein in the TI-complex resulted in a twofold increase in its immunogenicity. The formation of this vaccine construction was accompanied by significant conformational changes in the chimeric protein. Using HSP70 in the content of the chimeric protein represents an efficient means for presenting the main antigenic domain of the TBEV envelope protein to the immune system, whereas the incorporation of this chimeric protein into the TI-complex further contributes to the development of a stronger immune response against the TBEV infection.http://www.mdpi.com/2218-273X/8/3/82domain IIIenvelope proteinHSP70fusion proteinTBEVflavivirus vaccinedifferential scanning calorimetryintrinsic fluorescence |
| spellingShingle | Vasily Golotin Nina Sanina Ludmila Davydova Natalia Chopenko Andrey Mazeika Manuel Roig Valery Shnyrov Vladimir N. Uversky Eduard Kostetsky Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes Biomolecules domain III envelope protein HSP70 fusion protein TBEV flavivirus vaccine differential scanning calorimetry intrinsic fluorescence |
| title | Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes |
| title_full | Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes |
| title_fullStr | Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes |
| title_full_unstemmed | Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes |
| title_short | Recombinant Fusion Protein Joining E Protein Domain III of Tick-Borne Encephalitis Virus and HSP70 of Yersinia pseudotuberculosis as an Antigen for the TI-Complexes |
| title_sort | recombinant fusion protein joining e protein domain iii of tick borne encephalitis virus and hsp70 of yersinia pseudotuberculosis as an antigen for the ti complexes |
| topic | domain III envelope protein HSP70 fusion protein TBEV flavivirus vaccine differential scanning calorimetry intrinsic fluorescence |
| url | http://www.mdpi.com/2218-273X/8/3/82 |
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