Proximity hybridization regulated dual-mode ratiometric biosensor for estriol detection in pregnancy serum.

Sensitive and accurate determination of estriol level is vastly significant for the fetal growth and development. Herein, we constructed a dual-mode ratiometric biosensor for estriol assay combining the competitive immunoreaction, proximity hybridization with a two-step resonance energy transfer (RET) strategy. Estriol antibody and goat anti-rabbit antibody labeled DNA probes (Ab1-DNA1-Pt NPs and Ab2-DNA2) both hybridized with silver nanoclusters labeled DNA strands (H1-Ag NCs). Thus, the formed proximity hybridization enabled the occurrence of fluorescence RET (FL-RET, as the primary RET) between Ag NCs (donor) and Pt NPs (acceptor), quenching FL intensity of Ag NCs (FL off). When target estriol existed, the competitive reaction of Ab1-DNA1-Pt NPs with estriol and Ab2-DNA2 avoided the proximity hybridization. Then, the estriol-dependent H1-Ag NCs quenched electrochemiluminescence (ECL) emission of CdS quantum dots (CdS QDs, ECL off), generating ECL-RET (as the second RET). Consequently, according to the reverse changes of FL and ECL responses, this sensor realized the quantification of estriol from 1 to 100 ng/mL. Moreover, satisfactory results were achieved while testing estriol in pregnancy serum specimens, suggesting that the system is promising for potential application in samples analysis.