In order to create subunit vaccine against widespread and deadly viral disease - encephalitis, most feasible is the use of domain III of the tick-borne encephalitis virus (TBE) E protein, which includes the main virus neutralizing epitopes, as well as the adjacent C-terminal polypeptide chain, consisting of the stem and a hydrophobic anchor required for incorporation of the protein into the adjuvant nanocarrier of antigens - tubular immunostimulating complex (TI-complex) elaborated by us. It was used expression system of Escherichia coli and the cell-free expression system to obtain two fragments of E protein: domain III and domain III with the stem and the hydrophobic anchor. It was shown that hydrophobic anchor, which is toxic for bacteria, is destroyed after synthesis in the cell. However, part of E protein with hydrophobic anchor was successfully expressed in cell-free system. Immunization of mice by received antigens in the content of TI-complexes and without them showed no immunogenicity of domain III, while domain III with the stem and the anchor stimulated the production of antibodies by 2 times more effectively compared with the control. The incorporation of this recombinant protein in TI-complex resulted in further enhance of the antigen immunogenicity by 1.5 times. The results will be used to search for more effective forms of the C-terminal domain of E protein for optimal vaccine construction based on TI-complexes.
expression systems, subunit proteins, adjuvant nanocarrier of antigen
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