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: 2013-03-27
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 gold-based QCM chip into a mixed solution of bromoisobutylundecanethiol and trihydroxyethylundecanethiol with a total concentration of 1mM, in which bromoisobutylundecanethiol and The ratio of trihydroxyethylundecanethiol is 1:10, incubated at room temperature for 15 hours, then the QCM chip is taken out, rinsed with ethanol, and dried with nitrogen to obtain a QCM chip with a self-assembled layer modified on the surface; configuration For a mixed solution with a volume ratio of deionized water and methanol of 1:1, add 16mmol of 2,2'-bipyridine, 5mmol of oligoglycol methacrylate, and 5mmol of methacrylic acid to each liter of the mixed solution. Mix the hydroxyethyl ester and add 0.004mmol of copper chloride to obtain a light blue solution. Deoxidize the light blue solution for 30 minutes. Then add 0.004mmol ascorbic acid to each liter of the light blue solution after deoxygenation, and keep the nitrogen gas. Atmosphere, the red solution is obtained, ...

Embodiment 2

[0034] Step 1. Immerse the gold-based QCM chip into a mixed solution of bromoisobutylundecanethiol and trihydroxyethylundecanethiol with a total concentration of 10mM, in which bromoisobutylundecanethiol and The ratio of trihydroxyethylundecanethiol is 1:100, incubated at room temperature for 15 hours, then the QCM chip is taken out, rinsed with ethanol, and dried with nitrogen to obtain a QCM chip with a self-assembled layer modified on the surface; configuration For a mixed solution with a volume ratio of deionized water and methanol of 1:1, add 16mmol of 2,2'-bipyridine, 5mmol of oligoglycol methacrylate, and 5mmol of methacrylic acid to each liter of the mixed solution. Mix the hydroxyethyl ester and add 0.004mmol of copper chloride to obtain a light blue solution. Deoxygenate the light blue solution for 20 minutes. Then add 0.004mmol ascorbic acid to each liter of the light blue solution after deoxygenation, and keep the nitrogen gas. Atmosphere, the red solution is obtain...

Embodiment 3

[0041] Step 1. Immerse the gold-based QCM chip into a mixed solution of bromoisobutylundecanethiol and trihydroxyethylundecanethiol with a total concentration of 5mM, in which bromoisobutylundecanethiol and The ratio of trihydroxyethylundecanethiol is 1:50, incubated at room temperature for 15 hours, then the QCM chip is taken out, rinsed with ethanol, and dried with nitrogen to obtain a QCM chip with a self-assembled layer modified on the surface; configuration For a mixed solution with a volume ratio of deionized water and methanol of 1:1, add 16mmol of 2,2'-bipyridine, 5mmol of oligoglycol methacrylate, and 5mmol of methacrylic acid to each liter of the mixed solution. Mix the hydroxyethyl ester and add 0.004mmol of copper chloride to obtain a light blue solution. Deoxygenate the light blue solution for 10 minutes. Then add 0.004mmol ascorbic acid to each liter of the light blue solution after deoxygenation, and keep the nitrogen gas. Atmosphere, the red solution is obtained...

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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 specifically relates to a method for screening telomerase inhibitors by using a quartz crystal microbalance. Background technique [0002] Telomeres are repeated 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, while the length of somatic telomeres is much shorter than that of germ cells. In cell culture experiments, it was also observed that the length of telomeres decreased with the increase of the number of divisions, and the length of telomeres was inversely proportional to the degree of cell differentiation. The shortening of telomeres occurs every time a cell replicates, which is accompanied by shortening of telomeres. When telomeres are shortened to a certain extent, the stability of cell chromosomes will decrease, and the cells will gradually die. The number of tim...

Claims

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Application Information

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Patent Type & Authority Patents(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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