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<p>Ixodes ticks are vectors of many infectious human and animal pathogens. During the process of biting, the tick secretes specific proteins produced in its salivary glands. These proteins play key roles in various biological processes, including modulation of the host immune response, pathogen transmission, blood processing, and tick reproduction and development. Among these proteins, serpins have received special attention and are considered as promising candidates for the development of vaccines and methods to control infectious diseases transmitted by ticks. The aim of the work is to obtain and characterize in vitro recombinant RCL domain of serpin Ipis of ticks <i>Ixodes persulcatus</i>. PCR product was cloned into PCR 2.1 and pET-200 D/TOPO plasmids. Transfection was performed on chemically competent <i>E. coli</i> One Shot TOP10 and <i>E.&#xa0;coli</i> BL21(DE3) cells. The protein was purified from impurities by nickel chelate chromatography using Ni-NTA-Superflow resin. To analyze the immunogenicity of the obtained protein, mice of BALB/c line were immunized. The obtained sera of laboratory animals were tested by enzyme immunoassay. The structures of tick serpins were constructed by homology modeling using the AlphaFold server (<a href="https://www.alphafold.com/">https://www.alphafold.com/</a>). The recombinant RCL domain of serpin RCL of Ipis serpin of ticks <i>Ixodes persulcatus</i> was obtained and characterized in this study. The immunogenicity of the recombinant protein was evaluated by immunization of laboratory animals and the specific activity of the obtained sera was assessed by ELISA and immunoblot methods. 3D models of tick serpins were constructed. Thus, in this study, the recombinant RCL domain of serpin Ipis of <i>Ixodes persulcatus</i> ticks was obtained and characterized. After additional studies, the obtained recombinant RCL domain of serpin Ipis is planned to be used as mono- and multi-antigenic protein vaccines.</p>

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Production and Characterization In Vitro of Recombinant RCL Domain of Serpin Ipis of Ixodes persulcatus Ticks

  • 期刊论文

  • 发表日期：

    2025年9月1日

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  • K. V. Karelina,
  • E. V. Protopopova,
  • S. E. Pashkovskii,
  • V. A. Ternovoi,
  • R. B. Bayandin

摘要

Ixodes ticks are vectors of many infectious human and animal pathogens. During the process of biting, the tick secretes specific proteins produced in its salivary glands. These proteins play key roles in various biological processes, including modulation of the host immune response, pathogen transmission, blood processing, and tick reproduction and development. Among these proteins, serpins have received special attention and are considered as promising candidates for the development of vaccines and methods to control infectious diseases transmitted by ticks. The aim of the work is to obtain and characterize in vitro recombinant RCL domain of serpin Ipis of ticks Ixodes persulcatus. PCR product was cloned into PCR 2.1 and pET-200 D/TOPO plasmids. Transfection was performed on chemically competent E. coli One Shot TOP10 and E. coli BL21(DE3) cells. The protein was purified from impurities by nickel chelate chromatography using Ni-NTA-Superflow resin. To analyze the immunogenicity of the obtained protein, mice of BALB/c line were immunized. The obtained sera of laboratory animals were tested by enzyme immunoassay. The structures of tick serpins were constructed by homology modeling using the AlphaFold server (https://www.alphafold.com/). The recombinant RCL domain of serpin RCL of Ipis serpin of ticks Ixodes persulcatus was obtained and characterized in this study. The immunogenicity of the recombinant protein was evaluated by immunization of laboratory animals and the specific activity of the obtained sera was assessed by ELISA and immunoblot methods. 3D models of tick serpins were constructed. Thus, in this study, the recombinant RCL domain of serpin Ipis of Ixodes persulcatus ticks was obtained and characterized. After additional studies, the obtained recombinant RCL domain of serpin Ipis is planned to be used as mono- and multi-antigenic protein vaccines.