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Thermosensitive hydrogel electrochemiluminescence aptasensor, preparation method and application

An aptamer sensor, thermosensitive hydrogel technology, applied in chemiluminescence/bioluminescence, electrochemical variables of materials, analysis by chemical reaction of materials, etc. Accuracy and other issues, to achieve the effect of good selectivity, high sensitivity, and avoidance of interference

Active Publication Date: 2022-04-15
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These analysis methods include colorimetry, fluorescence, electrochemical methods, etc., but most of them have the limitations of high background signal, complicated operation, and inaccurate detection.

Method used

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  • Thermosensitive hydrogel electrochemiluminescence aptasensor, preparation method and application
  • Thermosensitive hydrogel electrochemiluminescence aptasensor, preparation method and application
  • Thermosensitive hydrogel electrochemiluminescence aptasensor, preparation method and application

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preparation example Construction

[0072] The preparation method of thermosensitive hydrogel material of the present invention comprises the following steps:

[0073] S1: Weigh 0.04g of N-isopropylacrylamide solid and 0.002g of N,N-methyldieneacrylamide solid in 1mL of distilled water, ultrasonically dissolve for 5-10 minutes to obtain N-isopropylacrylamide and N , N-methylenebisacrylamide mixed monomer solution;

[0074] S2: Weigh 19 mg ruthenium (II) (4-vinyl-4-methyl-2,2-pyridine) bis(2,2-pyridine) bis(hexafluorophosphoric acid) (relative molecular mass: 927.07) and dissolve In 10mL of acetone:distilled water mixed solvent with a volume of 1:1, ultrasonically dissolve for 1-3 minutes to obtain ruthenium(II)(4-vinyl-4-methyl-2,2-pyridine)bis(2, 2-bis(pyridine) bis(hexafluorophosphoric acid) solution, which is subsequently diluted according to experimental needs;

[0075] S3: Dissolve 2.0OD Apt-1 and 2.0OD Apt-2 sequences in 0.1M PBS buffer solution with pH=7.4 to obtain Apt-1 buffer solution with a concentr...

Embodiment 1

[0087] The preparation method of the dual-response electrochemiluminescence aptasensor based on thermosensitive hydrogel comprises the following steps:

[0088] S1: Add 10 μL of 100 μM Apt2 buffer solution to 10 μL of 10 mM TCEP solution, activate at room temperature for 1 hour, centrifuge at 8000 rpm for 10 minutes, and then disperse the precipitate in 10 μL of distilled water for later use to prepare activated Apt2 buffer solution;

[0089] S2: Add 10 μL of the Apt-2 buffer solution activated in step S1 to the 200 μL gold rod solution prepared above, and then stir and incubate at room temperature at 200 r / min for 10 h; then add 42 μL of 3M NaCl solution to the mixed solution for modification, and the modified After the gold rod solution was aged for 12 hours, it was centrifuged with 12000rpm ultrapure water for 15min, and the centrifuged precipitate was centrifuged with 12000rpm ultrapure water for 15min, and then centrifuged with 12000rpm ultrapure water for 15min. Store in...

Embodiment 2

[0103] Optimization of conditions for the detection of isocarbophos by a dual-response electrochemiluminescent aptasensor based on a thermosensitive hydrogel:

[0104] According to Example 1, a dual-response electrochemiluminescence aptasensor based on thermosensitive hydrogel was immersed in 5 mL of electrolyte containing different concentrations of tripropylamine (TPA) in pH=7.4 PBS buffer solution, and the photomultiplier tube was set at high pressure. It is 350V, and the scanning voltage range is 0.2V to 1.4V. After 808nm laser irradiation, electrochemiluminescence detection is performed. Change the concentration of tripropylamine in the electrolyte to be 2mM, 3mM, 4mM, 5mM, 6mM and 7mM respectively, and detect the electrochemiluminescence signal, the results are as follows Figure 8 As shown in A, when the concentration of TPA is 5mM, ΔECL responds best; TPA is a co-reactant of the ruthenium luminophore, reaching the optimal concentration can obtain the maximum ECL emissi...

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Abstract

The invention provides a dual-response electrochemiluminescence aptamer sensor based on thermosensitive hydrogel, a preparation method and application thereof. The aptamer chain Apt-1 at the 5' end of the bond is used as the recognition unit, and the gold rod-Apt-2 recognition element is added, and the plasmon resonance effect of the gold nanorod can enhance the ECL intensity of ruthenium terpyridine. The specific binding of the two split aptamers reduces the distance between the luminophore and the gold rod-Apt-2, which reduces the ECL signal. Moreover, the plasmonic photothermal effect of gold nanorods can satisfy the hydrogel phase transition condition, leading to the collapse of the hydrogel to further promote the RET effect. Furthermore, the linear relationship between the intensity of the signal change and the concentration of ammonium phosphine was constructed, and the sensitive detection of ammonium phosphine was realized.

Description

technical field [0001] The invention belongs to the technical field of electrochemical sensors, and in particular relates to a thermosensitive hydrogel-based dual-response electrochemiluminescence aptamer sensor, a preparation method and an application thereof, which can realize sensitive detection of organophosphorus pesticide isacarbophos. Background technique [0002] Typical thermosensitive microgels based on polyalkylacrylamide derivatives such as poly(N-isopropylacrylamide) (pNIPAM) have been widely used in recent years. These hydrogel particles swell in water below the so-called volume phase transition temperature (VPTT), and shrink above this temperature due to a change in the polymer's solvent affinity. Due to their high hydration state, they are generally biocompatible. Furthermore, their soft nature makes them particularly interesting for sensing and actuation, while their porous nature makes them attractive for the encapsulation and release of molecules. In vie...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/30G01N27/327G01N21/76G01N33/53
CPCG01N27/305G01N27/308G01N27/3278G01N21/763G01N21/76G01N33/53
Inventor 王广凤孙家慧
Owner ANHUI NORMAL UNIV
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