Graphical abstract Highlights • A ratiometric ECL biosensor was developed for the analysis of ovarian cancer marker. • A catalytically promoted ECL sensing platform based on NiFe 2 O 4 nanotubes was designed for ratiometric ECL strategy. • Envision complex was introduced into the ratiometric system to enhance the ECL emission process of two luminophors. • The developed ratiometric ECL biosensor exhibited excellent analytical performance in the detection of HE4. Abstract 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, NiFe 2 O 4 nanotubes (NiFe 2 O 4 NTs) with charming properties could act as catalytically promoted sensing platform. This NiFe 2 O 4 NTs based platform could trigger an intense ECL emission of DBAE for the reason that NiFe 2 O 4 NTs not only achieved the high loading of Envision-Ab 1 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 H 2 O 2 decomposition to generate superoxide (O 2 −) 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. [ABSTRACT FROM AUTHOR]