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Method for screening telomerase inhibitor with quartz crystal microbalance

A technology of telomerase inhibitors and quartz crystals, which is applied in the direction of material absorption weighing, measuring devices, instruments, etc., to save manpower, reduce false positive problems, and resist non-specific adsorption.

Inactive Publication Date: 2012-03-28
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But this special structure also inhibits the PCR process of the traditional method

Method used

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  • Method for screening telomerase inhibitor with quartz crystal microbalance
  • Method for screening telomerase inhibitor with quartz crystal microbalance
  • Method for screening telomerase inhibitor with quartz crystal microbalance

Examples

Experimental program
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Embodiment 1

[0027] Step 1, immerse the QCM chip of gold base into the mixed solution of bromoisobutylundecaptan and trihydroxyethyl undecaptan with a total concentration of 1mM, wherein bromoisobutylundecaptan and The ratio of trihydroxyethylundecaptanthiol is 1:10, incubate at room temperature for 15 hours, then take out the QCM chip, rinse it with ethanol, and dry it with nitrogen to obtain a QCM chip with a self-assembly layer modified on the surface; configuration Deionized water and methanol volume ratio are 1: 1 mixed solution, add 16mmol 2,2'-dipyridyl, 5mmol oligopolyethylene glycol methacrylate, 5mmol methacrylic acid-2- Hydroxyethyl esters were mixed, and 0.004 mmol of copper chloride was added to obtain a light blue solution, and the light blue solution was deoxygenated for 30 minutes, and then 0.004 mmol of ascorbic acid was added to each liter of deoxygenated light blue solution, and nitrogen atmosphere, to obtain a red solution, the QCM chip surface modified with a self-asse...

Embodiment 2

[0034] Step 1, immerse the QCM chip of gold base into the mixed solution of bromoisobutylundecaptan and trihydroxyethyl undecaptan with a total concentration of 10mM, wherein bromoisobutylundecaptan and The ratio of trihydroxyethylundecaptanthiol was 1:100, incubated at room temperature for 15 hours, then the QCM chip was taken out, rinsed with ethanol, and then dried with nitrogen to obtain a QCM chip with a self-assembly layer modified on the surface; configuration Deionized water and methanol volume ratio are 1: 1 mixed solution, add 16mmol 2,2'-dipyridyl, 5mmol oligopolyethylene glycol methacrylate, 5mmol methacrylic acid-2- Hydroxyethyl esters were mixed, and 0.004 mmol of copper chloride was added to obtain a light blue solution, and the light blue solution was deoxygenated for 20 minutes, and then 0.004 mmol of ascorbic acid was added to each liter of the deoxygenated light blue solution, and nitrogen was kept atmosphere, to obtain a red solution, the QCM chip surface m...

Embodiment 3

[0041] Step 1, immerse the QCM chip of gold substrate into the mixed solution of bromoisobutylundecaptan and trihydroxyethyl undecaptan with a total concentration of 5mM, wherein bromoisobutylundecaptan and The ratio of trihydroxyethylundecaptanthiol was 1:50, incubated at room temperature for 15 hours, then the QCM chip was taken out, rinsed with ethanol, and then dried with nitrogen to obtain a QCM chip with a self-assembly layer modified on the surface; configuration Deionized water and methanol volume ratio are 1: 1 mixed solution, add 16mmol 2,2'-dipyridyl, 5mmol oligopolyethylene glycol methacrylate, 5mmol methacrylic acid-2- Hydroxyethyl esters were mixed, and 0.004 mmol of copper chloride was added to obtain a light blue solution, and the light blue solution was deoxygenated for 10 minutes, and then 0.004 mmol of ascorbic acid was added to each liter of the deoxygenated light blue solution, and nitrogen gas was kept atmosphere, to obtain a red solution, the QCM chip su...

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Abstract

The invention discloses a method for screening a telomerase inhibitor with a quartz crystal microbalance. The method comprises the following steps: I, performing carboxyl-functionalizing of a chip surface; II, activating the chip surface; III, fixing a telomerase primer on the chip surface; IV, putting a chip on which the primer is fixed into a reactor of the quartz crystal microbalance, introducing a working solution into the reactor, reading the monitored frequency F1, then introducing a control solution, reading the monitored frequency F2, and calculating a difference (delta)F between F1 and F2; and V, putting the chip on which the primer is fixed into the reactor of the quartz crystal microbalance, introducing a working solution into the reactor, reading the monitored frequency F1', then introducing a liquid under test, reading the monitored frequency F2', calculating a difference (delta)F' between F1' and F2', and if F1' / (delta)F'<=50%, determining that the liquid under test contains the telomerase inhibitor. In the method disclosed by the invention, the quartz crystal microbalance is adopted as a detection tool, and the method has the advantages of no mark, real-time detection and the like.

Description

technical field [0001] The invention belongs to the technical field of drug screening, and in particular relates to a method for screening telomerase inhibitors by using a quartz crystal microbalance. Background technique [0002] Telomeres are repetitive sequences of DNA at the ends of chromosomes. In the 1990s, some researchers proposed that the average length of telomeres in the human body becomes shorter with age, and the telomere length of somatic cells is much shorter than that of germ cells. In cell culture experiments, it was also observed that the length of telomere shortened with the number of divisions increasing, and the length of telomere was inversely proportional to the degree of cell differentiation. Telomere shortening occurs every time a cell replicates and is accompanied by shortening of the telomere. When the telomere is shortened to a certain extent, the stability of the cell chromosome will be reduced, and the cell will gradually die. The number of c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N5/00G01N5/02
Inventor 马宏伟朱志强汪杰
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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