A SERS-enhanced controllable chain-like assembly for detection of thrombin

A technology of thrombin and assembly, applied in measuring devices, material stimulation analysis, instruments, etc., can solve the problems of antibody stability being easily affected by the environment, long antibody preparation cycle, and affecting the sensitivity and specificity of detection results, etc., to achieve Strong Raman signal enhancement effect, quantitative detection, and strong Raman enhancement effect

Active Publication Date: 2020-08-04
嘉兴千纯生物科技有限公司
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Problems solved by technology

[0005] Technical problem to be solved: The widely used immunological detection method for protein detection relies on the use of antibodies, and the preparation cycle of antibodies is long, and the stability of antibodies is easily affected by the environment, which further affects the sensitivity and specificity of detection results

Method used

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  • A SERS-enhanced controllable chain-like assembly for detection of thrombin
  • A SERS-enhanced controllable chain-like assembly for detection of thrombin

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Embodiment 1

[0029] A method for detecting thrombin by a SERS-enhanced controllable chain assembly, comprising the steps of:

[0030] (1) Synthesis of Au@Ag nanoparticles and Ag nanoparticles

[0031] Add 96mL of ultrapure water into a clean three-necked flask, then add 2.5% of chloroauric acid with a mass concentration of 0.4%, stir magnetically and heat to boil, add 1.5mL of trisodium citrate with a mass concentration of 1% after 10min, and the solution changes from After colorless to wine red, stop heating and continue stirring for 15 min to obtain 20nm gold nanoparticles. Take more than 5 mL of synthesized gold nanoparticles, add 200 μL of 1% PVP solution, 100 μL of 0.1 M ascorbic acid solution, and then add 70 μL of 0.1 mM silver nitrate solution, mix well, and shake at room temperature for 1 h in the dark That is, 50nm Au@Ag nanoparticles with 20nm Au core and 15nm thick Ag shell were obtained.

[0032]Add 20mL of ultrapure water, 5mL of 1% PVP, 0.5mL of 0.01mol / L sodium borohydrid...

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Abstract

The invention relates to a method for detecting thrombin by virtue of an SERS enhanced controllable chain-shaped assembly, belonging to the field of nano-sensing detection. According to the method, DNA molecules of two aptamers of the thrombin are respectively modified to the surfaces of an Au@ nano-particle and an Ag nano-particle, an alternate-arrayed chain-shaped structure is formed between thetwo nano-particles by controlling the DNA modification amounts of the surfaces of the nano-particles through the combination between assembly molecules and the thrombin when the thrombin exists, anda relatively strong Raman signal enhancement effect is achieved; and when the content of the thrombin is higher, the chain structure is longer, and the Raman signal strength is stronger, and the quantitative determination of the thrombin can be realized according to the corresponding relation between the content of the thrombin and the Raman signal strength. The method is simple in operation and high in sensitivity, and the thrombin in a serum sample can be detected; and by utilizing the nano-structure assembly, an important theoretical foundation is provided for detection of other target objects taking the aptamers as recognition molecules.

Description

technical field [0001] The invention belongs to the field of nanometer sensing and detection, and in particular relates to a method for detecting thrombin with a controllable chain assembly enhanced by SERS. Background technique [0002] Protein is the main carrier of life activities and the main executor of biological functions. Almost all physiological and pathological processes can cause corresponding changes in proteins. Therefore, protein detection is widely used in basic life science research, human health evaluation, disease diagnosis, medicine and food. It has high application value in analysis and biosafety monitoring, and the detection of disease-related proteins in the field of biomedical diagnosis is a very important task. [0003] At present, the commonly used methods for protein detection are immunoassay methods based on the specific recognition of antigens and antibodies, mainly including Western Blot, enzyme-linked immunosorbent adsorption (ELISA), affinity c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 杨蕾
Owner 嘉兴千纯生物科技有限公司
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