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Cytochrome (cyt ) plays a critical role in mitochondrial respiratory chain, whose absence is detrimental to electron transport and reduce adenosine triphosphate. For ultrasensitive detection of cyt , sheet-like covalent organic frameworks (COFs) were prepared by orderly accumulation of 1,3,5-benzenetricarboxaldehyde (BTA) and -phenylenediamine (PDA), and further grafted with -(4-aminobutyl)--ethylisoluminol (ABEI) - an electrochemiluminescence (ECL) emitter. Specifically, the morphology and structure of the COFs-ABEI were mainly characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD) analysis, and X-ray photoelectron spectroscopy (XPS). In parallel, the optical properties of the emitter were certified by UV-vis absorbance spectroscopy, Fourier infrared spectroscopy (FTIR), fluorescence (FL), and ECL measurements, showing 2.25-time enhanced ECL efficiency over pure ABEI, coupled by illustrating the interfacial electron transport mechanism. On the above foundation, a label-free "signal off" ECL biosensor was constructed by virtue of the specific immune recognition between the aptamer of the target cyt with its capture DNA (cDNA) anchored on the biosensing platform, exhibiting a wider linear range of 1.00 fg mL-0.10 ng mL ( = 0.998) and a lower limit of detection (LOD) down to 0.73 fg mL. This work offers some constructive guidelines for sensitive bioassays of disease-related biomarkers in the clinical field.
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