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Preparation method and application of photoelectrochemical biosensor based on iridium complex

A biosensor, iridium complex technology, applied in chemical instruments and methods, organic chemistry, scientific instruments, etc., can solve the problems of light damage of biological systems, excitation wavelength is limited to ultraviolet region, weak absorption, etc., and achieve high specificity Analysis of the effect of detection

Active Publication Date: 2019-07-23
王登之 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the bottleneck that restricts the further application of iridium complexes in the field of PEC lies in their weak absorption in the visible light region, and the maximum excitation wavelength is limited to the ultraviolet region, which is easy to cause photodamage to biological systems

Method used

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  • Preparation method and application of photoelectrochemical biosensor based on iridium complex
  • Preparation method and application of photoelectrochemical biosensor based on iridium complex
  • Preparation method and application of photoelectrochemical biosensor based on iridium complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of iridium complexes

[0032] Weigh IrCl 3 ·3H 2 O solid (0.35g, 1mmol) was dissolved in water (3mL), stirred to fully dissolve, added coumarin-6 (0.88g, 2.5mmol) and ethylene glycol ether (10mL), and the reaction mixture was placed under a nitrogen atmosphere React at 120°C for 27h, stop the reaction and cool to room temperature. Filtered, washed with absolute ethanol, evaporated to remove the solvent under reduced pressure, purified by silica gel column chromatography (100-200 mesh, eluent: dichloromethane / methanol=10:1), and then dried in vacuo to constant weight to obtain orange yellow Solid chlorine-bridged intermediate (0.58 g, 0.3 mmol), yield 60%. Weigh the obtained chlorine-bridged intermediate (0.185g, 0.1mmol), 2,2'-bipyridine-4,4'-dicarboxylic acid (0.06g, 0.25mmol), excess Na 2 CO 3 , was added to ethylene glycol ether (15 mL). Heated to reflux for 24 h under the protection of nitrogen, and cooled to room temperature. Remove the solvent b...

Embodiment 2

[0034] Preparation of Iridium Complex Colloidal Gold Nanoprobes

[0035] (1) Preparation of nano-gold: Wash the glass instruments, magnets and jars storing gold nanoparticles required in the preparation process with secondary water, soak them overnight with aqua regia, and then rinse with a large amount of ultrapure water Until the pH value is neutral, dry it for later use. Chlorauric acid HAuCl 4 (1.0mmol / L, 100mL) was stirred and heated to boiling in a cleaned one-necked flask, then trisodium citrate (38.8mmol / L, 10mL) was quickly added to the above solution, and the solution continued to react for 10 minutes. Turn into dark wine red, continue to reflux for 15 minutes, stop heating, naturally cool to room temperature while stirring, and store in the refrigerator at 4°C in the dark.

[0036] (2) Activation of the carboxyl group of the iridium complex: Dissolve the iridium complex (0.064g, 0.05mmol) in 10mL of anhydrous DMF, add EDC (0.058g, 0.3mmol), NHS (0.035g, 0.3mmol), ...

Embodiment 3

[0039] Based on the iridium complex gold nanoprobe prepared in Example 2, a photoelectrochemical biosensor was prepared for the detection of thrombin.

[0040] (1) Preparation of the working electrode: The ITO electrode was thoroughly cleaned with acetone, ethanol, and ultrapure water in sequence, and then fully dried under a nitrogen atmosphere. Soak the ITO electrode in 1mL ultrapure water, 0.2mL NH 4 OH (30%), 0.2mL H 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 APTES (5% (V / V)) , soak overnight at room temperature. Fully wash with ethanol to remove APTES that is not firmly adsorbed, fully dry in a nitrogen atmosphere, and then place it in an oven at 110 ° C for 1 h. Subsequently, it was soaked in the prepared colloidal gold solution overnight, thoroughly washed with ultrapure water, and drie...

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Abstract

The invention discloses a preparation method of a photoelectrochemical biosensor based on a cyclometalated iridium complex and application of the photoelectrochemical biosensor to thrombin detection.The biosensor uses the visible light-excited iridium complex as a photoelectrically active material, and the iridium complex is located on colloidal gold to prepare a gold nanometer signal probe. Capturing hairpin DNAs are immobilized on a modified ITO electrode to prepare a working electrode of the biosensor. A thrombin recognition system includes a specific recognition hairpin HP1 and an adjuvant hairpin HP2; in the presence of thrombin, a large number of secondary targets T-DNAs are released under the action of the excision enzyme ExoIII and hybridize with the capturing hairpin DNAs on theworking electrode so as to capture the gold nanometer signal probe, thereby realizing response to photocurrent signals; thrombin with different concentrations allows the amount of the signal probe connected to the working electrode to be different, so photocurrent intensities are different, and thus, quantitative detection of thrombin is realized. The method has the advantages of high sensitivityand good selectivity.

Description

technical field [0001] The invention relates to a preparation method of a photoelectrochemical biosensor based on a ring metal iridium complex and its application in thrombin detection, and belongs to the technical field of photoelectric functional materials and biological analysis. Background technique [0002] Biomarkers, such as microRNA, proteins, enzymes, etc., are a class of markers related to cell growth, proliferation, and disease occurrence that have been proposed in recent years with the development of immunology, molecular biology, and genomics technologies. , Play an important role in early disease diagnosis, prevention, drug target determination, drug response and other aspects, especially in the prevention and control of chronic and complex diseases such as tumors, cardiovascular diseases, and diabetes. As an important biomarker, thrombin can regulate physiological and pathological processes such as inflammation, anemia and wound healing speed, and its concentr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N21/63C07F15/00
Inventor 李春香卢维森庞萌萌周晓明朱明
Owner 王登之
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