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

IF 3.5 1区农林科学 Q1 VETERINARY SCIENCES

Veterinary Research Pub Date : 2025-08-14 DOI:10.1186/s13567-025-01602-z

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

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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.

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牛IP-10特异性单克隆抗体的制备与鉴定。

由于相对缺乏有效的试剂，对牛体内趋化因子的研究受到阻碍。牛IP-10是一种重要的炎症趋化因子，也是由牛分枝杆菌感染引起的一种重要的经济疾病（牛结核病）的诊断生物标志物。目前还没有针对牛IP-10的单克隆抗体。本研究的目的是利用杂交瘤技术制备检测牛IP-10的新型单克隆抗体。经交叉克隆抑制分析发现，5个单抗具有高度的自抑制作用。一个单抗（7C2）与一个商业多克隆抗体偶联，形成夹心ELISA。经重组牛IP-10检测，与现有的基于多克隆抗体的ELISA相比，该ELISA具有更强的线性范围。使用7C2的酶联免疫吸附试验可检测牛支原体感染动物样品中的牛特异性抗原IP-10。7C2单抗也能检测胞浆内IP-10。这些新的单克隆抗体将有助于阐明IP-10在健康、疾病和疫苗接种背景下的牛免疫研究中的作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Veterinary Research 农林科学-兽医学

CiteScore

7.00

自引率

4.50%

发文量

审稿时长

3 months

期刊介绍： Veterinary Research is an open access journal that publishes high quality and novel research and review articles focusing on all aspects of infectious diseases and host-pathogen interaction in animals.