Signal-off/on electrogenerated chemiluminescence deoxyribosensors for assay of early lung cancer biomarker (NAP2) based on target-caused DNA charge transfer.

Two novel electrochemiluminescence (ECL) deoxyribosensors are designed for assay of early lung cancer biomarker (NAP2) using the DNA three-way junction (DNA-TWJ) inserted NAP2 binding aptamer between two double-helical stems and labeled with ruthenium (II) complex (Ru) (NBAT-Ru) taken as molecular recognition element. The signal-off ECL deoxyribosensor was fabricated by covalently coupling the 5'-NH-(CH)-NBAT-Ru to glassy carbon electrode surface modified with 4-aminobenzoic acid (4-ABA). After combining NAP2 and NBAT-Ru, the changed conformation of NBAT-Ru altered the distance between Ru complex and electrode, resulting in a low ECL signal. The signal-on deoxyribosensor was fabricated by self-assembling the 5'-SH-(CH)-NBAT-Ru onto the Au electrode. The introduction of NAP2 triggered the conformational change in the aptamer domain, which induces the interhelical stacking of the two double-helical stems of NBAT-Ru. This stacking constitutes "electrical contact," which promotes transmission of electron-holes through the stems of NBAT-Ru, and produces high ECL intensity. Both deoxyribosensors show high sensitivity and selectivity. The biosensors have been successfully applied to clinical plasma detection. The approaches we describe represent unique principles based on DNA-TWJ inserted target special binding domain as molecular recognition element and different immobilization types for the fabrication of biosensors, which are greatly promising for the detection of protein, metal ions, bacteria, and cells.