Recombinant antigens in serological diagnostics of transboundary and emerging bovine infections

Introduction. Transboundary and emerging infections of cattle and small ruminants, such as peste des petits ruminants, Schmallenberg virus infection, etc., pose a serious animal health and economic threat in the context of developing globalization. Given the current geopolitical situation, the need for modern domestically produced diagnostic systems is particularly acute. Such systems can be developed using genetic engineering methods.

Objective. Analysis of domestic and foreign publications on the production of recombinant proteins of pathogens of transboundary and emerging infections of cattle and small ruminants. Creation of genetic constructs based on the processed data for further development of diagnostic tools, in particular ELISA test systems.

Materials and methods. Using bioinformatics tools, codon composition of the sequences encoding the nucleocapsid proteins of peste des petits ruminants virus (PPRV) and Schmallenberg virus (SBV) was analyzed and optimized. The optimized gene fragments were synthesized de novo and cloned into the pET-32b(+) expression vector. Successful insertion of the target sequence into the vector was confirmed by polymerase chain reaction and restriction analysis.

Results. Information on ELISA test systems developed on the basis of recombinant antigens for the diagnosis of peste des petits ruminants and Schmallenberg virus infection is presented. The main technological aspects of obtaining recombinant antigens for their further use in a diagnostic system factored in the biological features of a particular pathogen are highlighted. Our proprietary methodology for creating protein expression vectors for the pathogens of the diseases under review is additionally described.

Conclusion. The most promising recombinant antigens for use in ELISA test systems designed to detect antibodies against PPRV and SBV are full-length and truncated virion nucleocapsid proteins. Furthermore, the biophysical properties and antigenic structure of these proteins enable their production in Escherichia coli.  It should be noted that production of significant amounts of functional proteins in soluble form may require their expression as part of fusion proteins with tags enhancing solubility and facilitating correct folding.

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