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Molecularly imprinted intelligent gel grating, preparation method thereof and thrombin detection method

A technology of molecularly imprinted and smart gels, applied in molecularly imprinted smart gel gratings and their preparation, in the field of thrombin detection based on the molecularly imprinted smart gel gratings, can solve problems such as large errors and unreliable accuracy, Achieve the effects of improved recombination ability, sensitive and convenient detection, and high-sensitivity sensing detection

Pending Publication Date: 2021-11-05
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Spivak's research group used thrombin as a template to prepare aptamer-functionalized hydrogels using molecular imprinting technology. Using the specific recognition ability of aptamers for thrombin, they directly measured the volume change of the hydrogel to realize the recognition of thrombin. detection, but this method relies on manual measurement of gel volume changes, resulting in large errors, and the accuracy of detection needs to be improved

Method used

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  • Molecularly imprinted intelligent gel grating, preparation method thereof and thrombin detection method
  • Molecularly imprinted intelligent gel grating, preparation method thereof and thrombin detection method
  • Molecularly imprinted intelligent gel grating, preparation method thereof and thrombin detection method

Examples

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Embodiment 1

[0069] In this example, the molecularly imprinted smart gel grating was prepared, and the schematic diagram of its preparation process and thrombin response principle is shown in figure 1 As shown, the steps are as follows:

[0070] (1) Silanization of the substrate

[0071] Place the quartz glass sheet in concentrated sulfuric acid solution for 15 minutes to remove inorganic impurities on the surface of the quartz glass sheet, then place it in deionized water for 15 minutes and clean it with deionized water. Place the cleaned glass piece in acetone for 15 minutes to remove organic impurities on the surface of the glass piece, then place it in deionized water for 15 minutes and clean it with deionized water, and place the cleaned quartz glass piece in deionized water for later use . The purpose of this step is to keep hydrocarbon contamination to a minimum while ensuring that the quartz glass sheet remains fully wettable when immersed in a solution containing the silane coup...

Embodiment 2

[0083] In this example, to test the chemical composition of the molecularly imprinted smart gel grating, the steps are as follows:

[0084] (1) Measure the absorbance value of the DNA (A1 and A2) standard solution using a UV-visible spectrophotometer, and draw the DNA concentration-absorbance standard according to the content of DNA in the standard solution and its absorbance value at a wavelength of 257nm at ultraviolet light curves, such as image 3 As shown in a) figure, wherein the abscissa DNA concentration represents the ratio of the DNA test content to the theoretical addition. The linear regression equation of the standard curve is y=0.0021x-0.0005, the correlation coefficient R 2 It is 0.9984, indicating that the ratio of the DNA content to the theoretical added value follows Beer's law in the range of 0-14%, that is, the concentration of the light-absorbing substance DNA is proportional to its absorbance.

[0085] Therefore, by measuring the absorbance value of the...

Embodiment 3

[0088] In this example, the morphology characteristics of the molecularly imprinted smart gel grating prepared in Example 1 were characterized.

[0089] 1. The optical photo of the gel grating prepared in embodiment 1 is as Figure 4 As shown in Figure a, after peeling off the grating template, a gel grating with the same size as the gel grating template is obtained, and the size of the gel grating is 18mm×12mm. It can also be seen from the optical pictures that under natural light, a very obvious diffraction phenomenon occurs on the surface of the gel grating, that is, stripes of different colors can be observed on the surface of the gel grating.

[0090] The scanning electron microscope (SEM) photograph of the gel grating of embodiment 1 is as Figure 4 As shown in Figure b, it can be seen from the figure that the gel grating has a large-area regular microstructure, which ensures the stability and repeatability of the test results. At the same time, the gel grating has a m...

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Abstract

The invention provides a molecular imprinting intelligent gel grating and a preparation method thereof, the gel grating is composed of a substrate and parallel gel strips on the substrate, gaps between the gel strips are slits of the gel grating, and the gel strips are composed of thrombin response type molecular imprinting gel; the thrombin response type molecularly imprinted gel is formed by eluting thrombin in a gel network of a gel containing a thrombin imprinted compound. The invention further provides a thrombin detection method based on the molecular imprinting intelligent gel grating. The thrombin detection sensitivity and convenience can be improved, the detection limit of the thrombin detection method is as low as 10 <-12 > mol / L, thrombin at the concentration level of 10<-12> - 10<-7> mol / L can be detected, and the thrombin detection method is particularly suitable for ultrasensitive quantitative detection of thrombin in urine.

Description

technical field [0001] The invention belongs to the field of biomarker detection, and relates to a molecularly imprinted smart gel grating and a preparation method thereof, and a thrombin detection method based on the molecularly imprinted smart gel grating. Background technique [0002] Thrombin is a multifunctional serine protease. As the main effector protein in the blood coagulation cascade, it can promote the transformation of soluble fibrinogen in plasma into insoluble fibrin, and realize rapid hemostasis. At the same time, thrombin also acts as a hormone to regulate platelet aggregation, endothelial cell activation and other important responses in biological blood vessels. More importantly, recent reports indicate that thrombin can also be used as a biomarker in urine to detect disease signals such as glomerulonephritis, hepatitis, lupus, diabetes, and cancer. Therefore, it is of great significance to realize the sensitive detection of thrombin. [0003] For the det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/47G01N33/573
CPCG01N21/4788G01N33/573G01N2333/974
Inventor 褚良银赵佳佳汪伟蔡泉威谢锐巨晓洁刘壮
Owner SICHUAN UNIV