The exosome is a promising biomarker carrying many kinds of membrane proteins with huge heterogeneity, so the sensitive and multiplex analysis of exosomes is very significant for disease diagnosis and exploration of their biological functions. Herein, we propose an efficient method for highly sensitive detection and heterogeneity identification of exosomes based on the design and fabrication of an aptamer-coated liposome complex coupled with terminal deoxynucleotidyl transferase (TdT)-mediated polymerization. Specifically, in the presence of target exosomes, the aptamers immobilized on the surface of 1,2-dioleoyl-3-trimethylammonium-propane liposomes prefer to bind with exosomal membrane proteins due to the high affinity. The resulting aptamer-exosome complex will be accessible to TdT to switch on the polymerization reaction for signal amplification, achieving highly sensitive detection of exosomes. Furthermore, the proposed method can be employed to profile different exosomal membrane proteins by making use of a cluster of corresponding aptamers and obtain a fingerprint map of various cancer cell-derived exosomes. Thus, our approach may provide a highly sensitive and robust strategy for the identification of exosome heterogeneity with advantages of being label-free and having no separation, potentially enabling the precise subpopulation of exosomes with practical value in clinical applications.
Keywords: G-quadruplex; TdT; aptamer-coated liposome; exosome profiling; heterogeneity.