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Modified electrodes, combined products and their electrochemiluminescent biosensors and applications

A combined product and modified electrode technology, which is applied in the field of biosensors, can solve problems such as measurement errors, lack of reproducibility, and insufficient system stability, and achieve excellent signal amplification capabilities, good photostability, and easy surface functionalization.

Active Publication Date: 2020-05-22
SOUTHWEST UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The third object of the present invention is to provide an electrochemiluminescent biosensor assembled from the above-mentioned combination product, the electrochemiluminescent biosensor is an ECL biosensor without a co-reaction reagent, combined with a bipedal DNA walker (walker) dual The amplification strategy can overcome the shortcomings of the existing ECL biosensor assays in the presence of co-reaction reagents, such as insufficient system stability, measurement errors, and lack of reproducibility in detection, while having ideal ECL signal response values ​​and Excellent ECL luminous efficiency

Method used

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  • Modified electrodes, combined products and their electrochemiluminescent biosensors and applications
  • Modified electrodes, combined products and their electrochemiluminescent biosensors and applications
  • Modified electrodes, combined products and their electrochemiluminescent biosensors and applications

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0083] Experimental example 1 Synthesis of PFBT-COOH dots

[0084] First, PFBT and PSMA were dissolved in THF to prepare concentrations of 1.0 mg mL -1 stock solution. Then, 4.0 mL of the prepared PFBT and 800 μL of PSMA were mixed and sonicated to form a homogeneous mixed solution. Subsequently, 10 mL of double distilled water was injected into the above solution and stirred overnight. Finally, THF was removed by partial vacuum evaporation to obtain PFBT-COOH dots, which were stored in the dark.

Embodiment 1

[0085] The preparation of embodiment 1 biosensor

[0086] Fabrication of a bipedal DNA walker

[0087] Synthesis of a bipedal DNA walker by target-catalyzed hairpin assembly (CHA).

[0088] Heat the DNA hairpin H1 and DNA hairpin H2 to 95°C, keep the reaction for 5min, and then cool slowly to room temperature to form a hairpin structure;

[0089] Mix 10 μL of different concentrations of target miRNA-155 and 5 μL of 10 μmol L -1 Add DNA hairpin H1 to 5 μL 10 μmol L -1 DNA hairpin H2; then, the mixture was incubated at 37°C for 90min. Get a bipedal DNA walker.

[0090] Construction of biosensors

[0091] figure 1 The step-by-step construction and assembly process of the ECL biosensor is demonstrated.

[0092] a) The glassy carbon electrode (GCE, Φ=4.0mm) was polished with 0.30 and 0.05 μm alumina respectively, and then ultrasonically cleaned with ethanol and double distilled water in sequence;

[0093] b) Apply 10 μL of double-distilled water dispersion of PFBT-COOH do...

experiment example 2

[0098] Experimental example 2 Characterization of biped DNA walker

[0099] The bipedal DNA walker synthesized by target-catalyzed hairpin assembly was analyzed by polyacrylamide gel electrophoresis (PAGE).

[0100] The prepared DNA chain was dropped into the groove of a newly prepared polyacrylamide gel (16%), and then electrophoresed in 1×TBE buffer solution with a potential of 120V for 120 min. After staining with ethidium bromide (EB), images were taken with a biogel imaging system. Such as figure 2 As shown, the missing band in lane 1 corresponds to miRNA-155 (1.0 μmol L -1 ), because it has few nucleotide bases and does not stain easily. Lane 2 and lane 3 respectively show the corresponding H2 (1.0μmol·L -1 ) and H1 (1.0μmol·L -1 ) of different bands. Two independent bands appeared in lane 4 (1.0 μmol·L -1 H2 and 1.0 μmol L -1 H1), indicating that H2 and H1 can coexist stably in solution. As expected, when miRNA-155 (1.0 μmol·L -1 ), H1 (1.0 μmol L -1 ) and H...

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Abstract

The invention relates to a modified electrode, a combined product, an electrochemiluminescent biosensor and application. The modified electrode comprises an electrode and a luminous body modified on the electrode, and the luminous body mainly includes carboxyl functionalized poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1',3 }-thiadiazole)] points. The electrochemiluminescent biosensor hasno coreaction reagent, and can overcome the defects including insufficient system stability, measuring errors and lack of reproducibility in detection in present ECL biosensor determination with a coreaction reagent, a double signal amplification electrochemiluminescence strategy without the coreaction reagent is formed, the ECL signal response value and ECL light emitting efficiency are high, the sensitivity, specific selectivity and stability are high, and a new method is provided for nucleic acid detection.

Description

technical field [0001] The invention relates to the field of biosensors, in particular to modified electrodes, combined products and electrochemiluminescent biosensors and applications thereof. Background technique [0002] MicroRNAs (miRNAs) are endogenous, non-protein-coding, short (usually about 19-25 bases) and single-stranded RNAs that have been shown to be involved in several biological processes such as hematopoiesis, cell proliferation and apoptosis important regulators in . Studies have shown that miRNAs can be used as ideal biomarker candidates, and the abnormal expression of miRNAs is closely related to the occurrence of various cancers. Therefore, there is an urgent need to develop highly sensitive, highly specific and reliable techniques to detect miRNAs. [0003] Electrochemiluminescence (ECL) not only has the advantages of electrochemical methods, such as simplicity and stability of equipment, but also has a wider dynamic range and higher sensitivity than co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/30G01N27/327G01N27/416
CPCG01N27/305G01N27/308G01N27/3275G01N27/416
Inventor 陈时洪李芹谭兴容
Owner SOUTHWEST UNIV
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