Herein, a delicate ratiometric electrochemiluminescence biosensor was designed for the detection of human epididymis protein 4 (HE4) based on 2-(Dibutylamino)ethanol (DBAE) and lucigenin as anodic and cathodic ECL emitter, respectively. Initially, NiFe2O4 nanotubes (NiFe2O4 NTs) with charming properties could act as catalytically promoted sensing platform. This NiFe2O4 NTs based platform could trigger an intense ECL emission of DBAE for the reason that NiFe2O4 NTs not only achieved the high loading of Envision-Ab1 complex but effectively catalyzed the oxygen evolution reaction (OER) to yield oxygen. Moreover, the Envision complex, a polymeric material containing enrich horseradish peroxidase (HRP) could further accelerate the H2O2 decomposition to generate superoxide (O2−) for the enhancement of the ECL emission process of two luminophors. It’s noteworthy that hexagonal boron nitride nanosheets (h-BN) as the nanolabels, was first illustrated to be capable of immobilizing a large quantity of lucigenin and retaining intense and stable ECL emission of lucigenin in the alkaline condition. Benefiting from the above merits, a new ECL ratiometric sensing mode was established for the sensitive determination of HE4 in a range from 10 fg/mL to 10 ng/mL with the detection limit of 3.3 fg/mL. The strategy exhibited broad prospect in early ovarian cancer diagnosis and blazed a new path for other tumors screening.