Telomerase activity testing system based on SPR technology

A testing system, telomerase technology, applied in the direction of measuring devices, material analysis through optical means, instruments, etc., can solve the problems of complex measurement methods, long time, high cost, etc., to reduce costs, simplify the experimental process, improve safety effect

Inactive Publication Date: 2017-01-04
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These measurement methods are complex, time-consuming and costly

Method used

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  • Telomerase activity testing system based on SPR technology
  • Telomerase activity testing system based on SPR technology
  • Telomerase activity testing system based on SPR technology

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0035] 1) Prepare the test kit, which contains 10mmol / L Tris-HCl buffer solution, the pH of the solution is 7.3, and the buffer solution also contains deoxymononucleotide (dNTP) in equimolar proportions for the synthesis of DNA, the total concentration of dNTP 8mmol / L;

[0036] 2) Construction of the chip: After plating a layer of silver film with a thickness of 43nm on the glass slide, place it in a 5 μmol / L solution of thiol-modified telomeric DNA and incubate at 0°C for 10 hours, and connect the telomere DNA to the silver via the thiol group. The membrane surface, wherein the DNA sequence is as follows: 5'-SH-AAA AAA AAA AAA TTA GGG TTA GGG TTA GGG TTA GGG. After the modification is completed, block with 1 mmol / L mercaptoethanol phosphate buffer for 2 hours, and finally vacuum-dry the prepared chip and store it under nitrogen protection. It can be stored for a long time and used directly in the test;

[0037] 3) Use: Put the modified chip into the SPR instrument, add the w...

example 2

[0039] 1) Prepare the kit, which contains 200mmol / L Tris-HCl buffer solution, the pH of the solution is 7.3, and the buffer solution also contains deoxygenated mononucleotide (dNTP) in equimolar proportions for the synthesis of DNA, the total concentration of dNTP 12mmol / L;

[0040] 2) Construction of the chip: After plating a layer of gold film with a thickness of 46 nm on the glass slide, place it in 15 μmol / L telomeric DNA solution modified with trithioadamantane derivatives and incubate at 10°C for 24 hours. The structural formula of alkane derivatives is as follows:

[0041]

[0042] Wherein, R represents the functional group of trithioadamantane derivatives,

[0043]The sequence of the telomere DNA is: 5'-X-AAA AAA AAA AAA TTA GGG TTA GGG TTA GGG TTA GGG, where X is a trithioadamantane derivative, and the telomere DNA is connected to the surface of the gold membrane through a trimercapto group. After the modification is completed, block with 3 mmol / L mercaptoethanol...

example 3

[0046] 1) Prepare the test kit, which contains 100mmol / L Tris-HCl buffer solution, the pH of the solution is 7.3, and the buffer solution also contains deoxygenated mononucleotide (dNTP) in equimolar proportions for the synthesis of DNA, the total concentration of dNTP 10mmol / L;

[0047] 2) Construction of the chip: After plating a layer of gold film with a thickness of 44nm on the glass slide, place it in 10 μmol / L solution of telomeric DNA modified with sulfhydryl groups and incubate at 4°C for 20 hours, and connect the telomeric DNA to gold via sulfhydryl groups. The membrane surface, wherein the DNA sequence is as follows: 5'-SH-AAA AAA AAA AAA TTA GGG TTA GGG TTA GGG TTA GGG. After the modification is completed, block with 2 mmol / L mercaptohexanol phosphate buffer for 4 hours, and finally vacuum-dry the prepared chip and store it under nitrogen protection. It can be stored for a long time and used directly in the test;

[0048] 3) Use: Put the modified chip into the SPR ...

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Abstract

The invention provides a telomerase activity testing system based on an SPR technology. The telomerase activity testing system comprises a kit containing a buffer solution of 10-200 mmol/L and dNTP of 8-12 mmol/L, and an SPR chip provided with a glass sheet, a metal coating film covering the glass sheet and a telomerase DNA connected to the coating film, wherein the pH of the buffer solution is 7.3. Signal sources such as radioactive isotope or fluorescent markers are not needed, the safety is improved, and the detection costs are greatly reduced. The influence of PCR experiments is eliminated, further detection is achieved, and the experiment process is simplified. Separation and detection conducted on an amplification product are not needed, the detection speed is greatly improved, the detection real-timeliness and repeatability can be achieved, and the experiment reliability is improved.

Description

technical field [0001] The invention relates to a testing system for telomerase activity. More specifically, the present invention relates to a telomerase activity testing system based on SPR technology. Background technique [0002] Telomerase (telomerase) is a kind of reverse transcriptase, which is an RNA ribose-protein that can use telomere DNA as a substrate to repeat DNA amplification at the end of telomere DNA at the end of a eukaryotic linear chromosome Complex. The amplification of telomere DNA by telomerase can make cells divide indefinitely without natural apoptosis, thus leading to the occurrence of cancer. At present, the overexpression of telomerase has been found in more than 85% of cancer cells, and there is a high correlation between telomerase and tumor cells. It may become a new target of anti-tumor drugs and an important marker for tumor diagnosis. Establishing an efficient, rapid and sensitive analytical method is of great significance for screening a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552
CPCG01N21/553
Inventor 陈名利尹焕才田晶晶陆一泓
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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