Proximity hybridization-triggered signal switch for homogeneous chemiluminescent bioanalysis

A proximity hybridization-triggered signal switch was presented for convenient homogeneous chemiluminescent detection of a wide range of affinity target biomolecules, such as oligonucleotides, protein biomarkers, and aptamer-recognized targets. The presence of the target promoted the formation of a proximate complex via the proximity hybridization of two help DNA strands or the DNA strands labeled to affinity ligands, which subsequently unfolded the self-reporting molecular beacon to switch on the chemiluminescence signal. The response could be further amplified with an in situ enzymatic recycling strategy for highly sensitive chemiluminescence detection. By using an antibody as the affinity ligand, this simple protocol could sensitively detect protein biomarker in a concentration range of 6 orders of magnitude with a detection limit down to 80 fM. With the use of an aptamer as the affinity ligand, a method for homogeneous chemiluminescent detection of thrombin was proposed. The one-step and wash-free assay showed good selectivity and required only 1 μL of sample. Its high sensitivity, acceptable accuracy, and satisfactory versatility of analytes led to various applications in bioanalysis.