Fluorescent oligonucleotide probe based on G-quadruplex scaffold for signal-on ultrasensitive protein assay

Abstract

This work reported the G-quadruplex structure of pyrene-labeled G-rich DNA probe and its application in the immunoglobulin E (IgE) detection, providing plausibly an insight into the biological function of human telomere. Based on the intermolecular G-quadruplex, a terminal-single-pyrene-labeled oligonucleotide signaling probe was developed and a novel protein assay strategy was proposed via combining specific DNA cleavage by S1 nuclease with target-recognizing aptamer. This assay platform not only circumvented the optimization of specific sites for reporter attachment and the pyrene monomer fluorescence quenching by flanking nucleotide bases but also presented a signal-on mechanism. Thus, ultrasensitive homogeneous detection of IgE was successfully conducted. A linear dynamic range of 4.72 × 10 −12 to 7.56 × 10 −9 m, a regression coefficient of 0.9941 and a detection limit of 9.45 × 10 −14 m were given. Additionally, a preliminary concept of the single-pyrene-labeled excimer fluorescence probes associated with G-quadruplex for screening biological markers was described. Importantly, the unexpected structural property of G-quadruplex discovered seems to provide valuable information to allow understanding of the structure and function of human telomere and exploring of useful structure-based anticancer drug.