Method for electrochemically detecting activation level of STAT 3 protein through DNA probe ligand and application

Abstract

The invention discloses a method for electromagnetically detecting the activation level of an STAT 3 protein through a DNA probe ligand and application. By use of STAT3 which is an important transcription factor and dynamic reversible and controllable mutual action between two natural ligands which are DNA and polypeptide, a DNA probe capable of specifically identifying a protein is designed; the polypeptide is used for controlling reversible combination and dissociation between the target protein STAT3 and the DNA probe; by stable coordination interaction between Zr<4+> and a phosphate group and a phosphorylated nano-graphene composite beacon, catalytic signal amplification is realized. According to the method, by introduction of the phosphorylated nano-graphene composite beacon serving as a signal amplification strategy, the phosphorylated nano-graphene composite beacon is stable and difficult to inactivate, thus finally realizing sensitive, efficient and stable detection for the activated STAT3 protein.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis and high-risk pregnancy detection in obstetrics, and relates to a method and application for electrochemically detecting STAT 3 protein activation level by using a DNA probe ligand. Background technique [0002] At present, the detection and analysis of protein activation level mainly rely on traditional immunological methods such as Western Blotting and ELISA; the detection of STAT 3 protein activation level is also mainly through these two methods. However, these classical methods relying on antigen-antibody recognition mechanisms have limitations to varying degrees, such as the highly complex chemical structure of antibodies, complicated preparation processes and high prices, cumbersome operations, time-consuming, and the need to contact radioactive substances. With the increasing interest in the specific detection of disease-related protein markers, in addition to immunoassays based on antig...

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Problems solved by technology

However, these classical methods relying on the antigen-antibody recognition mechanism have limitations to varying degrees, such as the highly complex chemical structure of the antibody, complicated preparation process and high price, cumbersome operation, time-consuming and need to contact radioactive substances, etc.

With the increasing interest in the specific detection of disease-related protein markers, in addition to immunoassays based on antigen-antibody binding, another type of protein biosensor based on chemically synthesized ligands of nucleic acid aptamers has also received great attention. The two complement each other, but the detection of the activation level of protein markers still faces new challenges in terms of sensitivity, selectivity, and response speed: First, the rigid one-to-one ratio between a single ligand such as an antibody and a target protein The binding effect can easily cause signal attenuation, which affects the further introduction of new signal amplification strategies; second, the "sandwich" detection system designed by using two different ligands to recognize different domains on the target protein is not universally applicable, and its application range is limited. Relatively limited, because not all target proteins can have a satisfactory "sandwich" binding site

Based on the shortcomings of existing protein analysis and detection technologies, such as static, low efficiency, and low spatiotemporal resolution, it is difficult to meet the needs of life science research and clinical medical testing.

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