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DNA aptamer modified bioelectrochemical sensor and preparation method thereof

A DNA aptamer, bioelectrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve problems such as high recurrence rate and low cure rate

Inactive Publication Date: 2010-09-08
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, in the existing treatment methods, the low cure rate and high recurrence rate are also problems in the process of cancer treatment

Method used

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  • DNA aptamer modified bioelectrochemical sensor and preparation method thereof
  • DNA aptamer modified bioelectrochemical sensor and preparation method thereof
  • DNA aptamer modified bioelectrochemical sensor and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Modification of gold electrodes

[0023] Before aptamer modification, the gold electrodes were sanded on fine sandpaper and then on Al 2 o 3 The powder was ground, and then ultrasonicated in absolute ethanol and ultrapure water for 5 minutes. In order to better remove impurities remaining on the surface of the gold electrode, place the electrode in 1M H 2 SO 4 In the 0-1.5V voltage range, the cyclic voltammetry (CV) peak sweep is performed, the sweep speed is set to 100mv / s, and the stability is reached in about 20 cycles. The electrodes were then blown dry in nitrogen for aptamer modification. The gold electrodes were respectively placed on the DNA aptamer chains containing sulfhydryl groups (5'-HS-(CH 2 ) 6 -ATCTA ACTGC TGCGC CGCCG GGAAA ATACT GTACG GTTAG A-3') in Eppendorf tubes containing HS-DNA aptamers in the concentration range of 0.05pM, 0.5pM, 5pM and 50pM, 10mM Tris-HCl, 1mM EDTA, 1M NaCl and 0.5 μM TCEP (pH 8.0), inverted for 16 hours. Afte...

Embodiment 2

[0024] Embodiment two: the cultivation and processing of cells

[0025] CCRF-CEM cells (human acute lymphoblastic leukemia T lymphocytes) and HL-60 cells (human myeloid leukemia cells) were purchased from Shanghai Institute of Cells, Chinese Academy of Sciences. Both cells were maintained at 37 °C, 5% CO 2 The culture medium was 1640 medium containing 10% fetal bovine serum, and the cells were collected when the cells grew to the end of the logarithmic growth phase for cell detection research. Count with a hemocytometer before detection, and the concentration of cells used each time is generally 10 5 about.

Embodiment 3

[0026] Example 3: Optimization of the concentration of the aptamer modified on the gold electrode

[0027] Cyclic voltammetry, the electrochemical probe used is 5mM K 3 [Fe(CN) 6 ] / K 4[Fe(CN) 6 ]. Electrochemical probe detection was performed on the gold electrodes numbered A, B, C, and D above, and the results were as follows: figure 1 . From the results of cyclic voltammetry, it can be observed that with the decreasing concentration of aptamer, K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] and the electrochemical transmission of the gold electrode surface becomes easier, so the signal is also increasing continuously, among which the B gold electrode surface and the K 3 [Fe(CN) 6 ] / K 4 [Fe(CN) 6 ] had the best electrochemical transfer, and a better peak shape was obtained. Afterwards, when the aptamer concentration was further reduced, the signal did not increase significantly.

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Abstract

The invention relates to a novel DNA aptamer modified bioelectrochemical sensor and a preparation method thereof. The novel bioelectrochemical sensor is of a three-electrode system. The invention is characterized in that in the three electrodes, the counter electrode is a platinum electrode, the reference electrode is a saturated calomel electrode, and the working electrode is a gold electrode; and a DNA aptamer chain is modified on the gold electrode. The novel DNA aptamer modified bioelectrochemical sensor combines the characteristics of the high specificity of the DNA aptamer and the high sensitivity of an electrochemical detection method and ensures that the early diagnosis of cancer cells becomes possible.

Description

technical field [0001] The invention relates to a novel bioelectrochemical sensor and a preparation method thereof, in particular to a novel bioelectrochemical sensor modified by a DNA aptamer and a preparation method thereof. Background of the invention [0002] Cancer is one of the diseases that seriously threaten human health in today's society. Among them, acute lymphoblastic leukemia is a particularly obvious threat to children's health. According to literature reports, nearly two-thirds of patients with acute lymphoblastic leukemia are children. Moreover, in the existing treatment methods, the low cure rate and high recurrence rate are also problems in the process of cancer treatment. However, if cancer can be diagnosed at an early stage, the patient's survival rate will be greatly improved, and studies have also proved that early diagnosis and timely treatment are one of the effective ways to improve the patient's survival rate. Therefore, it is extremely important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/327
Inventor 李根喜陈桂芳刘月星李婷陈震宇陈琳
Owner SHANGHAI UNIV
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