Development of a platform for producing recombinant protein components of epitope vaccines for the prevention of COVID-19статья
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Дата последнего поиска статьи во внешних источниках: 8 апреля 2022 г.
Аннотация:A new platform for creating anticoronavirus epitope vaccines has been developed. Two loop-like epitopes with lengths of 22 and 42 amino acid residues were selected from the receptor-binding motif of the Spike protein from the SARS-CoV2 virus that participate in a large number of proteinprotein interactions in the complexes with ACE2 an- neutralizing antibodies. Two types of hybrid proteins, including one of the two selected epitopes, were constructed. To fix conformation of the selected epitopes, an approach using protein scaffolds was used. The homologue of Rop protein from the Escherichia coli ColE1 plasmid containing helixturnhelix motif was used as an epitope scaffold for the convergence of C- and N-termini of the looplike epitopes. Loop epitopes were inserted into the turn region. The conformation was additionally fixed by a disulfide bond formed between the cysteine residues present within the epitopes. For the purpose of multimerization, either aldolase from Thermotoga maritima, which forms a trimer in solution, or alphahelical trimerizer of the Spike protein from SARS-CoV2, was attached to the epitopes incorporated into the Rop-like protein. To enable purification on the heparincontaining sorbents, a short fragment from the heparin-binding hemagglutinin of Mycobacterium tuberculosis was inserted at the C-terminus of the hybrid proteins. All the obtained proteins demonstrated high level of immunogenicity after triplicate parenteral administration to mice. Sera from the mice immunized with both aldolase-based hybrid proteins and the Spike protein SARSCoV2 trimerizer-based protein with a longer epitope interacted with both the inactivated SARS-CoV-2 virus and the Spike protein receptor-binding domain at high titers.