Design and production of recombinant multiepitope antigen to indicate antibodies against the virus arthritis-encephalitis of goats

To date, the only effective means of combating goat arthritis-encephalitis viruses (CAEV) includes its early diagnosis and slaughter of all animals in dysfunctional and suspected farms, since specific preventive measures have not been developed. Among the widely available tools for diagnostic studies, PCR indication of the CAEV genome and the detection of specific antibodies of the virus can be distinguished, which directly depends on the serological activity and specificity of the antigens used in diagnostic kits. This work is devoted to the development of a recombinant antigen for serological indication of antibodies against CAEV. As a result of bioinformatic analysis, a search was performed for amino acid sequences of virus proteins, an analysis of immunogenic epitopes of these proteins, a search for N-, O-glycosylation sites, signal peptide sites and transmembrane domains. Based on the data obtained, a recombinant multiepitope antigen was designed with the following amino acid sequence "mqktnepyedfaarlleaidaepipvgaeiipesmkylrgaksqgklneeaerwrrnnppppavraytygviempenyakvcqplvqiyrtlstptyqhhhhhh». The constructed amino acid sequence, after converting it to a nucleotide one, taking into account the codon optimization of protein production in Escherichia coli, was cloned into an expression construct based on the pET28a(+) vector. As a result of the design, the length of the created plasmid for the expression of the nucleotide sequence of the CAEV was 5527 bp. The protein developed and purified by affinity chromatography, containing amino acid sequences of immunogenic epitopes of the CAEV antigens, confirmed its functionality in indirect solid-phase enzyme immunoassay using blood sera from goats with previously confirmed immune status to this virus. The recombinant multiepitope antigen presented in this paper can be widely used as an integral component in the development of diagnostic test systems.

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