Sensitive and reliable detection of tumour markers is of great significance for early diagnosis and monitoring recurrence of cancers. Herein, a simple electrochemical immunosensor is developed with an integrated electrochemical probe on the sensing surface, which is able to sensitively and reagentlessly detect the breast cancer biomarker, human epidermal growth factor receptor 2 (ErbB2). Ferrocene (Fc) is chosen as the signal indicator and covalently grafted on cationic polyelectrolyte poly(ethylene imine) (Fc-PEI). The redox Fc-PEI could alternately assemble with carboxyl functionalized single-walled carbon nanotubes (SWNTs) on an indium tin oxide electrode through layer-by-layerelectrostatic assembly. After Anti-ErbB2 antibody is covalently immobilized onto the outermost SWNTs layer followed by blocking the electrode with bovine serum albumin, a sensing interface with recognitive probe and electrochemical probe is obtained. In the presence of ErbB2, the formed antigen-antibody complex makes a barrier to inhibit electro-transfer of inner Fc, leading to a decreased electrochemical response. Owing to the SWNTs-facilitated charge transfer and abundant surface-bound probes, the developed sensor demonstrates outstanding performance for reagentless detection of ErbB2 in terms of wide detection range (1.0-200.0 ng mL) and low detection limit (0.22 ng mL). The developed immunosensor also exhibits good selectivity, reproducibility and stability. Real analysis of ErbB2 in human serum samples is also demonstrated.