The extracellular vesicles show great potential as a noninvasive biomarker for the early detection of cancer.Hence,there is an urgent requirement to create biosensors that are time-saving,simple,and easily scalable in...The extracellular vesicles show great potential as a noninvasive biomarker for the early detection of cancer.Hence,there is an urgent requirement to create biosensors that are time-saving,simple,and easily scalable in order to accomplish rapid,sensitive,and quantitative detection of extracellular vesicles.In this study,we present a self-propelled DNA walker powered by endonuclease Nt.Bbv CI,which enables the development of a“signal on”sensing platform for the rapid and highly sensitive detection of extracellular vesicles.The DNA motor employed tracks made of streptavidin magnetic beads,which consisted of substrate strands labeled with fluorescein and motor strands locked by aptamers.The aptamer recognition of the target protein on extracellular vesicles unlocked the motor strand,initiating the DNA motor process.After replacing the optimal buffer solution containing the endonuclease Nt.BbvC I,the motor strands autonomously moved along the streptavidin magnetic beads track,continuously releasing fluorescent molecules and producing detectable fluorescence signals.Under optimal conditions,the detection range was from 2×10~4particles/mL to 2×10~9particles/mL,with a detection limit of 2.9×10~3particles/mL,demonstrating excellent selectivity.This method has demonstrated good selectivity in different tumorderived extracellular vesicles and performs well in complex biological samples.The ability to effectively analyze surface proteins of extracellular vesicles in a short period of time gives our DNA walker a tremendous potential for developing simple and cost-effective clinical diagnostic devices.展开更多
基金supported financially by the National Natural Science Foundation of China(NSFC,Nos.62071119 and 62075098)the National Key Research and Development Program of China(Nos.2017YFA0205301 and 2018YFC1602905)。
文摘The extracellular vesicles show great potential as a noninvasive biomarker for the early detection of cancer.Hence,there is an urgent requirement to create biosensors that are time-saving,simple,and easily scalable in order to accomplish rapid,sensitive,and quantitative detection of extracellular vesicles.In this study,we present a self-propelled DNA walker powered by endonuclease Nt.Bbv CI,which enables the development of a“signal on”sensing platform for the rapid and highly sensitive detection of extracellular vesicles.The DNA motor employed tracks made of streptavidin magnetic beads,which consisted of substrate strands labeled with fluorescein and motor strands locked by aptamers.The aptamer recognition of the target protein on extracellular vesicles unlocked the motor strand,initiating the DNA motor process.After replacing the optimal buffer solution containing the endonuclease Nt.BbvC I,the motor strands autonomously moved along the streptavidin magnetic beads track,continuously releasing fluorescent molecules and producing detectable fluorescence signals.Under optimal conditions,the detection range was from 2×10~4particles/mL to 2×10~9particles/mL,with a detection limit of 2.9×10~3particles/mL,demonstrating excellent selectivity.This method has demonstrated good selectivity in different tumorderived extracellular vesicles and performs well in complex biological samples.The ability to effectively analyze surface proteins of extracellular vesicles in a short period of time gives our DNA walker a tremendous potential for developing simple and cost-effective clinical diagnostic devices.
