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

## TL;DR

This study developed new monoclonal antibodies for bovine IP-10, a chemokine important for diagnosing bovine tuberculosis, and demonstrated their utility in detecting the protein in infected cattle.

## Contribution

The study introduces novel monoclonal antibodies for bovine IP-10 and validates their use in diagnostic assays.

## Key findings

- Five monoclonal antibodies were developed, with one (7C2) showing strong self-inhibition and utility in ELISA.
- The 7C2-based ELISA improved detection range and sensitivity compared to existing polyclonal antibody methods.
- The 7C2 antibody detected both native and intracytoplasmic IP-10 in infected cattle samples.

## 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.

The online version contains supplementary material available at 10.1186/s13567-025-01602-z.