Development and characterization of monoclonal antibodies specific for bovine IP-10
Abstract 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 bovi...
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| Format: | Article |
| Language: | English |
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BMC
2025-08-01
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| Series: | Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s13567-025-01602-z |
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| author | Hamza Khalid Michael Coad Inga Dry Catherine McGuinnes Lindsey A. Waddell Jayne C. Hope Zhiguang Wu |
| author_facet | Hamza Khalid Michael Coad Inga Dry Catherine McGuinnes Lindsey A. Waddell Jayne C. Hope Zhiguang Wu |
| author_sort | Hamza Khalid |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-caabe01b317d42aab9d672bd2068d7b8 |
| institution | Kabale University |
| issn | 1297-9716 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj-art-caabe01b317d42aab9d672bd2068d7b82025-08-20T03:46:29ZengBMCVeterinary Research1297-97162025-08-0156111110.1186/s13567-025-01602-zDevelopment and characterization of monoclonal antibodies specific for bovine IP-10Hamza Khalid0Michael Coad1Inga Dry2Catherine McGuinnes3Lindsey A. Waddell4Jayne C. Hope5Zhiguang Wu6Division of Immunology, The Roslin Institute, University of EdinburghAnimal and Plant Health AgencyDivision of Immunology, The Roslin Institute, University of EdinburghDivision of Immunology, The Roslin Institute, University of EdinburghDivision of Immunology, The Roslin Institute, University of EdinburghDivision of Immunology, The Roslin Institute, University of EdinburghDivision of Immunology, The Roslin Institute, University of EdinburghAbstract 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.https://doi.org/10.1186/s13567-025-01602-zIP-10intracellular staininghybridoma technologymonoclonal antibodybovine tuberculosis |
| spellingShingle | Hamza Khalid Michael Coad Inga Dry Catherine McGuinnes Lindsey A. Waddell Jayne C. Hope Zhiguang Wu Development and characterization of monoclonal antibodies specific for bovine IP-10 Veterinary Research IP-10 intracellular staining hybridoma technology monoclonal antibody bovine tuberculosis |
| title | Development and characterization of monoclonal antibodies specific for bovine IP-10 |
| title_full | Development and characterization of monoclonal antibodies specific for bovine IP-10 |
| title_fullStr | Development and characterization of monoclonal antibodies specific for bovine IP-10 |
| title_full_unstemmed | Development and characterization of monoclonal antibodies specific for bovine IP-10 |
| title_short | Development and characterization of monoclonal antibodies specific for bovine IP-10 |
| title_sort | development and characterization of monoclonal antibodies specific for bovine ip 10 |
| topic | IP-10 intracellular staining hybridoma technology monoclonal antibody bovine tuberculosis |
| url | https://doi.org/10.1186/s13567-025-01602-z |
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