Engineering of Chimeric Protein Based on E Protein Domain III of Tick- Borne Encephalitis Virus and OmpF Porin of Yersinia pseudotuberculosis

Abstract

The newly obtained chimeric antigen could be valuable for the development of the preventing tick-borne encephalitis subunit vaccines.

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

“…This observation indicated that the EIII protein, which is toxic to the E. coli host cells, obviously cannot be expressed separately from other proteins, which probably masks its toxic effects on producing cells. Therefore, we used the EIII protein produced using the continuous exchange cell-free system, as described earlier [ 8 ].…”

“…The plasmids 40HSP70/EIII and 40/EIII were obtained by ligation of the EIII gene into 40HSP70 and pET-40b(+), respectively, at the restriction sites of SacI and SalI. The resultant 40HSP70/EIII and 40/EIII plasmids were used for the expression of chimeric HSP70/EIII protein and the recombinant domain III of TBEV protein E. Since the EIII protein was not expressed separately due to its high toxicity for Escherichia coli cells, we used the EIII protein obtained using the cell-free continuous exchange system, as described previously [ 8 ]. All used primers are shown in Table 1 .…”

“…However, DIII is not immunogenic due to its low molecular weight (MW 16 kDa). One of the strategies for increasing the immunogenicity of this protein is the creation of chimeric (hybrid) recombinant proteins with specified properties and with decreased DIII toxicity for bacterial host cells [ 8 ]. Heat shock proteins (HSPs) serve as promising fusion partners due to their remarkable effects on the immune system [ 9 ].…”

Immunogenicity and Protective Activity of a Chimeric Protein Based on the Domain III of the Tick-Borne Encephalitis Virus E Protein and the OmpF Porin of Yersinia pseudotuberculosis Incorporated into the TI-Complex

“…To provide some clues to the structural basis of the efficient immunogenicity of the OmpF-EIII, we evaluated the intrinsic disorder predisposition of this chimeric protein using a comprehensive computational approach based on the utilization of several tools designed for characterization of various flavors of intrinsic disorder. As described earlier [ 10 ], the amino acid sequence of the chimeric OmpF-EIII protein includes 499 residues:…”

“…The domain III (DIII) of E protein is the main antigenic domain, which includes virus-specific epitopes recognized by neutralizing antibodies [ 8 , 9 ]. Therefore, we have constructed the chimeric protein OmpF-EIII based on the fusion of EIII, which consists of DIII and a stem of E protein, and porin OmpF of Gram-negative bacteria Yersinia pseudotuberculosis , for the further development of a subunit anti-TBE vaccine [ 10 ]. Membrane protein OmpF is necessary as a hydrophobic anchor, which allows incorporation of the fusion antigen into the lipid matrix of the nanoparticulate adjuvant and the antigen delivery system, such as the tubular immunostimulating complexes (TI-complexes) [ 5 , 11 ].…”