Nano-pore electrical sensor used for enzyme reaction detection

Abstract

The invention discloses a nano-pore electrical sensor used for enzyme reaction detection. The nano-pore electrical sensor used for enzyme reaction detection comprises a substrate layer, an insulationlayer, and a thin film which are overlapped from bottom to top successively; the center of the thin film is provided with a nanopore; the internal part of the nanopore is provided with a DNA probe through chemical modification, and a DNA tetrahedral structure capable of realizing complementary hybridization with the DNA probe; the DNA tetrahedral structure is combined with horseradish peroxidase labelled streptavidin through biotin, so that single horseradish peroxidase is immobilized in the nanopore; a reaction substrate is added to detect oxidation reaction product translocation events and realize real-time monitoring on enzyme reaction. The nano-pore electrical sensor used for enzyme reaction detection is capable of solving problems in the prior art that in enzyme reaction detection large amounts of reagent and plenty of time are consumed, and the enzyme immobilization method of enzyme in the nanopore is capable of providing research ideas for study on the molecular dynamics of thesingle enzyme in the nanopore.

Description

technical field [0001] The invention relates to a nanopore electrical sensor technology for enzyme reaction detection, belonging to the technical field of nanopore electrical sensors. Background technique [0002] Nanopore sensing is a widely-studied novel sensing and detection technology involving single-molecule random sensing, medical screening and diagnosis, etc. Nanopore sensing and detection technology originated from the principle of Coulter counting and cell ion channels. In nanopore sensing detection, nanopores or nanochannels (usually less than 100nm) are usually located on an insulating film, which divides the fluid pool into two to form two separate ion solution pools, that is, the cis-surface (Cis) and trans face (Trans). When a steady bias voltage is applied across these sides, a steady flow of ions from one side of the pore to the other is generated. When charged molecules pass through the nanopore, current blocking will occur, and the physical and chemical...

AI Technical Summary

Problems solved by technology

At present, most of the research on enzymes using nanopores is limited to detecting the conformational changes of enzyme molecules through the event of enzyme molecules passing through the holes, and immobilizing a large number of enzyme molecules in the pores to detect the occurrence of reactions. These methods consume a lot of reagents. , and can only conduct statistical analysis on a large number of enzyme molecules, and cannot realize the study of enzyme-catalyzed reactions at single-molecule resolution

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