Detecting The Secretion Of Exosomes From Individual Cells In Real Time By A Multifunctional Nanoelectrode-nanopore Nanopipette

Exosomes have received extensive attention for their fundamental roles in biosystem and as a novel biomarker for early detection and therapeutic applications.As a carrier of intercellular information,exosomes are involved in the occurrence,metastasis,drug resistance and immune escape of tumor cells,and play an important role in the spread of disease.A large number of circulating exosomes contained in body fluids also often carry cancer-related biological signal molecules.Therefore,sensitive exosome isolation and analysis technology is essential in clinical detection.We developed a new method based on glass nanoelectrodes to fabricate multifunctional nanoelectrode-nanopore nanopipettes(MNNPs)for the detection of exosomes.This study mainly focused on monitoring the exosomes of individual human cervical cancer cells(Hela cells)in real time while maintaining cell viability.The main research contents are as follows:1.Successfully prepared carbon electrode(CNEs)based MNNPs combined with resistance pulse sensing technology to detect exosomes extracted from Hela cells in real time.Analyzing the nanopore current signals generated by exosomes translocation events and detecting the average current blocking duration(?t).2.Modified CD63 antibody(anti-CD63)on the carbon electrode surface to optimize the MNNPs targeting of exosome.MNNPs with modified CD63 antibodies were fabricated by electrochemical methods,and translocation events induced by exosomes and liposomes were studied.3.Real-time monitoring the release of exosomes by individual living cells using MNNPs.Co-precipitation with Hela cells via exosome inhibitor GW4869,the number of current signals induced by exosome translocation behavior was significantly reduced.Furthermore,the non-modified MNNPs were used as control expriments and the targeting of modified antibodies was confirmed.