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<p>Research on chemokines in cattle is hampered by the relative lack of validated reagents. Bovine IP-10 is an important inflammatory chemokine and a promising diagnostic biomarker for an economically important disease (bovine tuberculosis) caused by infection of cattle with <i>Mycobacterium</i><i> bovis</i>. Currently no monoclonal antibodies are available for bovine IP-10. The goal of this study was to generate novel mAbs for detection of bovine IP-10 using hybridoma technology. Five mAbs were developed and cross clone inhibition analyses showed a high degree of self-inhibition among the mAbs. One mAb (7C2) was used in conjugation with a commercial polyclonal antibody to develop a sandwich ELISA. Upon testing with recombinant bovine IP-10, this ELISA showed an enhanced linear range compared to the currently available polyclonal antibody-based ELISA. The ELISA using 7C2 was shown to detect native antigen-specific bovine IP-10 in samples from <i>M. bovis</i> infected animals. The 7C2 mAb could also detect intracytoplasmic IP-10. These novel mAbs will be useful in elucidating roles for IP-10 in bovine immune studies in health, disease and vaccination contexts.</p>

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Development and characterization of monoclonal antibodies specific for bovine IP-10

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年8月14日

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  • Hamza Khalid,
  • Michael Coad,
  • Inga Dry,
  • Catherine McGuinnes,
  • Lindsey A. Waddell,
  • Jayne C. Hope,
  • Zhiguang Wu

摘要

Research on chemokines in cattle is hampered by the relative lack of validated reagents. Bovine IP-10 is an important inflammatory chemokine and a promising diagnostic biomarker for an economically important disease (bovine tuberculosis) caused by infection of cattle with Mycobacterium bovis. Currently no monoclonal antibodies are available for bovine IP-10. The goal of this study was to generate novel mAbs for detection of bovine IP-10 using hybridoma technology. Five mAbs were developed and cross clone inhibition analyses showed a high degree of self-inhibition among the mAbs. One mAb (7C2) was used in conjugation with a commercial polyclonal antibody to develop a sandwich ELISA. Upon testing with recombinant bovine IP-10, this ELISA showed an enhanced linear range compared to the currently available polyclonal antibody-based ELISA. The ELISA using 7C2 was shown to detect native antigen-specific bovine IP-10 in samples from M. bovis infected animals. The 7C2 mAb could also detect intracytoplasmic IP-10. These novel mAbs will be useful in elucidating roles for IP-10 in bovine immune studies in health, disease and vaccination contexts.