Monoclonal antibody-based indirect competitive ELISA for quantitative detection of Enterobacteriaceae siderophore enterobactin

•A monoclonal antibody (mAb) directed against Enterobacteriaceae siderophore enterobactin (Ent) was successfully generated for the first time with high affinity and specificity.•Based on the anti-Ent mAb, an indirect competitive ELISA (ic-ELISA) was developed for Ent quantification with high accuracy, repeatability, and reproducibility.•The ic-ELISA was able to quantitatively detect Ent produced in different bacterial cultures, laying a solid foundation for Ent-based diagnostics of gut health.