Preparation of rabbit monoclonal antibody against T-2 toxin and development of enzyme-linked immunosorbent assay in milk, feed and pork samples

• Developed RmAb targeting T-2 toxin with high specificity and affinity. • Optimized one-step ic-ELISA improved T-2 detection sensitivity in tissues. • New ic-ELISA method detects T-2 in under 1 h, ideal for rapid clinical testing. T-2 toxin (T-2) is a fungal toxin commonly found in contaminated grains. It can remain in animal-derived products after eating contaminated feed, and has potential teratogenic and carcinogenic effects. Previous T-2 toxin immunoassay detection methods often utilize mouse monoclonal antibodies (MmAbs). However, in comparison to MmAbs, rabbit monoclonal antibodies (RmAbs) demonstrate higher affinity and sensitivity. This study successfully prepared an RmAb targeting T-2 using single B cell clone technology, with an IC 50 of 0.089 ng/ml. By optimizing the working concentrations of the coating antigen and antibody, a one-step ic-ELISA method for detecting T-2 in milk, feed and Pork Samples was established based on RmAb, with a detection limit 0.32 to 1.42 µg/kg. This method reduces reaction time to under 1 h and achieves average recovery rates of 75.5–100.8 %, and the CV was less than 11.6 %, indicating high accuracy, repeatability, and reproducibility of the ic-ELISA. In conclusion, this study developed a highly specific and sensitive RmAb and established a one-step ic-ELISA based on this antibody for detecting T-2. The method proved to be reliable and rapid, providing a valuable reference for advancing T-2 residue detection techniques in foods.