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• PD-MWCNT nanocomposite containing CMC provided a large number of antibody binding sites. • An electrochemical immunosensor for zearalenone was developed by the U-disk portable potentiostat with smartphone. • zearalenone in feed ingredients and agricultural food was detected using a simple rapid portable immunoassay. Zearalenone (ZEN) has emerged as a global public health concern due to its significant pathogenicity and world-wide distribution in animal feed ingredients and agricultural food. In the study, we report on the design and development of a simple portable immunosensor based on polyamindoamine dendrimer (PD)-multi-walled carbon nanotube (MWCNT) nanocomposite containing carboxymethyl cellulose (CMC) for rapid electrochemical immunoassay of ZEN residual in feed ingredients and agricultural food using U-disk potentiostat with smartphone. A large number of ZEN monoclonal antibody was covalently immobilized onto the electrode surface of nanocomposite owing to abundant carboxyl groups of PD, MWCNT and CMC. Nanocomposite modified screen printed electrode was employed to fabricate a simple rapid portable immunosensor using U-disk potentiostat with smartphone. The electrochemical immunosensor presented a desirable linear relationship ranges from 0.001–100 ng mL−1 with a low detection limit of 0.15 pg mL−1 under the optimal conditions, which was consistent with the results of the immunosensor prepared with glassy carbon electrode on the traditional potentiostat with desktop computer. It is proposed as a simple and robust analytical tool for rapid detection of hazardous substances including ZEN in feed ingredients and agricultural food on the basis of the great performance of the proposed immunosensor. [ABSTRACT FROM AUTHOR]