Preparation method of photoelectric chemical biosensor for microRNA detection

A biosensor, photoelectrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of high electron-hole recombination rate, cadmium ion toxicity, poor photostability, etc., and achieve high sensitivity and stability good, specific effect

Active Publication Date: 2019-01-04
QINGDAO UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this type of material has a high electron-hole recombination rate, poor photostabi

Method used

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  • Preparation method of photoelectric chemical biosensor for microRNA detection
  • Preparation method of photoelectric chemical biosensor for microRNA detection
  • Preparation method of photoelectric chemical biosensor for microRNA detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Synthesis and characterization of cyclometalated iridium complex optoelectronic materials:

[0035] Weigh IrCl 3 ·3H 2 O solid (0.34g, 1mmol) was dissolved in water (5mL), stirred to fully dissolve, added coumarin-6 (0.77g, 2.2mmol) and ethylene glycol ether (15mL), and the reaction mixture was placed under a nitrogen atmosphere React at 120°C for 24h and cool to room temperature. Filter, wash with water, absolute ethanol and acetone to obtain a chlorine-bridged intermediate. Weigh the obtained chlorine-bridged intermediate (0.14 g, 0.08mmol), dipyrido[3,2-a:2',3'-c]phenazine (0.06g, 0.2mmol), and add to ethylenedi Alcohol ether (10mL). React at 120° C. for 15 h under nitrogen protection, and cool to room temperature. The solvent was removed by evaporation under reduced pressure and purified by column chromatography (200-300 mesh, eluent: dichloromethane / methanol=10 / 1) to obtain a dark red solid (0.061g, 0.046mmol, 58%). 1 H NMR (CDCl 3,500MHz)δ:9.69(d,J=9.0Hz,2H...

Embodiment 2

[0037] Preparation of ITO working electrode:

[0038] The ITO electrode was ultrasonically cleaned with acetone, ethanol, and ultrapure water in sequence, and then fully dried under a nitrogen atmosphere. Immerse the ITO electrode in 1mL ultrapure water, 0.2mLNH 4 OH (30%), 0.2mLH 2 o 2 (30%) in the mixed solution, take it out after soaking for 10min, continue to fully clean with ultrapure water, fully dry in nitrogen atmosphere, soak the above-mentioned treated ITO electrode in the ethanol solution of 3-aminopropyltrimethoxysiloxane Medium (5% (V / V)), 12 hours at room temperature. Fully wash with ethanol, fully dry in a nitrogen atmosphere, and then place it in an oven at 110° C. for 1 hour. Subsequently, it was soaked in the prepared colloidal gold solution and assembled for 12 hours, fully cleaned with ultrapure water, and dried under nitrogen. The capture hairpin HP1 was dissolved in SPSC buffer solution (1M NaCl, 50mM NaCl 2 HPO 4 , 10 mM tris(2-carboxyethyl)phosph...

Embodiment 3

[0040] Preparation of photoelectrochemical biosensors for microRNA detection:

[0041] 20 μL of different concentrations of microRNA-122b was dropped onto the ITO working electrode prepared in Example 2, incubated at 37° C. for 2 h, fully washed with Tris-HCl (0.01 M) buffer, and dried under nitrogen atmosphere. Then 20 μL of the mixture of hairpin HP2 and HP3 (1 μM) was added dropwise, incubated at 37°C for 2 hours, and after being fully washed with Tris-HCl (0.01M) buffer, 10 μL of the cyclometalated iridium complex prepared in Example 1 was added dropwise. Substance solution (solvent: DMF / PBS=1:20, 0.01 μM) was incubated for 40 minutes, and then washed with buffer solution to remove non-specifically adsorbed signal substances for the detection of photoelectric signals.

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Abstract

The invention discloses a preparation method of a cyclometalated iridium complex biosensor for microRNA detection. The preparation method includes: designing and synthesizing a cyclometalated iridiumcomplex insertable into DNA dual strands as a photoelectric material; fixing capture hairpin DNA to a modified ITO electrode to obtain a working electrode of the biosensor herein; if target microRNA exists, hybridizing with the capture hairpin DNA fixed on the working electrode to release its neck portion, and allowing the released neck portion to hybridize with another two hairpin DNAs to triggerhybridization chain reaction, so that a long DNA dual-strand polymer is formed on the surface of the working electrode. Massive cyclometalated iridium complexes can be inserted into the polymer herein so as to amplify signals, and therefore, high-sensitivity detection of microRNA is achieved. The preparation method has the advantages of high sensitivity, good specificity, good stability and the like.

Description

technical field [0001] The invention belongs to the field of photoelectric functional materials and biological analysis, in particular to a preparation method of a ring metal iridium complex biosensor for detecting microRNA. Background technique [0002] MicroRNAs are a class of endogenous non-coding single-stranded small RNA molecules, usually 19-23 nucleotides in length, which play an important role in cell proliferation, differentiation, apoptosis and other life processes. Studies have shown that the abnormal expression of microRNAs is closely related to many diseases, such as tumors, diabetes, heart disease, neurological diseases, etc., and has become a new type of tumor-specific biomarkers. Therefore, the realization of highly sensitive detection of microRNA is of great significance for the study of its biological function and early diagnosis of cancer. However, due to the short sequence of microRNAs, high sequence homology, and low expression level, it is extremely di...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/26
CPCG01N27/26G01N27/3278
Inventor 王墨泽蔡月圆李忠成
Owner QINGDAO UNIV OF SCI & TECH
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