Preparation method of high-throughput liquid crystal detection platform for screening enzyme inhibitor by inducing aptamer to release through enzyme catalysis
A detection platform and aptamer technology, which is applied in the field of analysis and detection, can solve problems such as restricting development, and achieve good effects, high sensitivity, and good selectivity
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[0053] A second aspect of the present invention provides a method for preparing a high-throughput liquid crystal biosensor for screening enzyme inhibitors by enzymatically inducing aptamer release, comprising:
[0054] Pretreatment of glass substrates;
[0055] Add liquid crystal molecules to the chloroform solution containing surfactant (OTAB), and then remove the chloroform with nitrogen to obtain a mixture of liquid crystal and OTAB, which is added dropwise to the surface of the copper mesh;
[0056]The inhibitory drug to be tested is mixed with substrate-converting specific enzyme, small molecule substrate, and aptamer DNA successively, and added dropwise on the above-mentioned copper grid to obtain the product.
[0057] The invention adopts the copper mesh method to observe the optical phenomenon of the liquid crystal, and has high sensitivity, short time consumption and simple preparation method.
[0058] A third aspect of the present invention provides a method for hig...
Embodiment 1
[0110] A high-throughput liquid crystal detection platform for screening xanthinease inhibitors using enzyme-catalyzed aptamer release to detect the enzyme inhibition efficiency of febuxin
[0111] The liquid crystal sensor used was prepared as follows:
[0112] a. Soak the glass sheet in the washing solution at 75°C for 30 minutes, then wash it with a large amount of ultrapure water, methanol, and ethanol, blow it dry with nitrogen, and dry it in an oven at 120°C for 10 to 20 hours.
[0113] b. At room temperature, soak the glass sheet in DMOAP with a volume fraction of 1% for 20 minutes, then wash it with ultrapure water, and dry it with nitrogen to obtain a glass substrate.
[0114] Several copper meshes were pipetted onto the resulting glass substrates.
[0115] Dissolve OTAB in chloroform to make a chloroform solution with a concentration of 1 mMOTAB.
[0116]c. Mix 10 μL of 5CB molecules with 90 μL of 1 mM OTAB solution, vortex for 30 seconds to obtain a final concentr...
Embodiment 2
[0124] Example 2: Synthesis of various xanthinease inhibitors and screening of effective drugs using a high-throughput liquid crystal detection platform for screening xanthinease inhibitors by inducing aptamer release through enzyme catalysis
[0125] The preparation of liquid crystal sensor is identical with embodiment 1
[0126] S1. Pre-mix the compound to be detected with xanthine oxidase and incubate for 30 minutes; then incubate with xanthine for 30 minutes, and then incubate with xanthine aptamer DNA for 60 minutes.
[0127] S2. Pipette 40 μL of the prepared mixture onto a liquid crystal sensor, and observe using a polarizing microscope.
[0128] The synthesized xanthine enzyme inhibitory drugs are all benzotriazole compounds and their derivatives. Through the above-mentioned liquid crystal detection platform, three suitable enzyme inhibitor drugs can be screened out with high throughput and effectively. Each structure is shown in Figure 5.
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