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Biomolecules 2018, 8(3), 82; https://doi.org/10.3390/biom8030082

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

1
Department of Biochemistry, Microbiology and Biotechnology, Far Eastern Federal University, Sukhanov St., 8, Vladivostok 690091, Russia
2
Laboratory of Marine Natural Compounds Chemistry, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, FEB RAS, Prospect 100 let Vladivostoku, 159, Vladivostok 690022, Russia
3
Departamento de Química Física, Facultad de Ciencias Químicas, Universidad de Salamanca, Plaza de los Caìdos s/n, 37008 Salamanca, Spain
4
Departamento de Bioquímica y Biología Molecular, Facultad de Biología, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain
5
Department 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, USA
6
Laboratory of New methods in Biology, Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino 142290, Moscow Region, Russia
*
Author to whom correspondence should be addressed.
Received: 4 July 2018 / Revised: 15 August 2018 / Accepted: 20 August 2018 / Published: 25 August 2018
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Abstract

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. View Full-Text
Keywords: domain III; envelope protein; HSP70; fusion protein; TBEV; flavivirus vaccine; differential scanning calorimetry; intrinsic fluorescence domain III; envelope protein; HSP70; fusion protein; TBEV; flavivirus vaccine; differential scanning calorimetry; intrinsic fluorescence
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Golotin, V.; Sanina, N.; Davydova, L.; Chopenko, N.; Mazeika, A.; Roig, M.; Shnyrov, V.; Uversky, V.N.; Kostetsky, E. 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 2018, 8, 82.

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