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Photo-thermal release signal enhanced type thyroglobulin electrochemiluminescence immunosensor

A thyroglobulin and signal-releasing technology, applied in electrochemical variables of materials, scientific instruments, instruments, etc., can solve the problems of inability to directly functionalize immune complexes, limit the application of TPA solid-phase luminescence, etc., and achieve highly sensitive detection, The effect of enhancing the ECL signal

Active Publication Date: 2019-10-08
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bipyridyl ruthenium Ru(bpy) 3 2+ Classic co-reaction reagents such as tripropylamine (TPA) have no free amino groups on the surface, so they cannot directly functionalize immune complexes, which greatly limits the application of TPA in ECL solid-phase luminescence

Method used

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  • Photo-thermal release signal enhanced type thyroglobulin electrochemiluminescence immunosensor
  • Photo-thermal release signal enhanced type thyroglobulin electrochemiluminescence immunosensor
  • Photo-thermal release signal enhanced type thyroglobulin electrochemiluminescence immunosensor

Examples

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Effect test

Embodiment 1

[0026] A method for preparing an electrochemiluminescent immunosensor based on photothermal release signal enhancement:

[0027] (1) Pretreatment of the glassy carbon electrode: the glassy carbon electrode is first mechanically polished and polished on the suede covered with alumina powder, washed with secondary water to remove the residual powder on the surface, and then moved into an ultrasonic water bath for cleaning until it is cleaned, and finally Wash thoroughly with ethanol, dilute acid and water;

[0028] (2) Mix 50 μL of 10 mg / mL mesoporous silica (MPS) aqueous solution with 50 μL of 10 mM bipyridyl ruthenium [Ru(bpy) 3 2+ ] The solution was shaken and absorbed for 6 h, and the mixture was centrifuged for 10 min to obtain the precipitate, which was washed and diluted with 100 μL of ultrapure water; then, 10 μL of 3-aminopropyltriethoxysilane (APTES) was added to the solution to functionalize After washing with ultrapure water for 12 h, 10 μL of 5 wt.% glutaraldehyde...

Embodiment 2

[0034] Preparation of mesoporous silica (MPS) used in Example 1: Weigh 0.5 g of cetyltrimethylammonium bromide (CTAB) and dissolve it in 240 mL of ultrapure water, add 3.5 mL of 2.0 M sodium hydroxide Solution, the temperature of the mixed solution was raised to 80 °C under full stirring, and the reaction was incubated for 30 min, and 2.5 mL of tetraethyl orthosilicate (TEOS) was slowly added dropwise, and then the stirring reaction was continued at 80 °C for 2 h, and the mixture was centrifuged for 10 min , the obtained precipitate was washed three times with ultrapure water and ethanol, and a white solid was obtained after vacuum drying; finally, 0.5 g of the obtained solid was added to a mixed solution of 50 mL of methanol and 2.5 mL of 37wt.% concentrated hydrochloric acid, heated to reflux for 16 h, The filtered precipitate was washed three times with ultrapure water and methanol, and dried under vacuum at 60 °C to remove the residual solvent in the pores to obtain mesopor...

Embodiment 3

[0037] A thyroglobulin electrochemiluminescent immunosensor based on photothermal release signal enhancement to detect thyroglobulin, the steps are as follows:

[0038] (1) Using a three-electrode system for measurement, the MPS-Ru(bpy) prepared in Example 1 3 2+ / TEPA / TG / Ab / MB modified electrode is the working electrode, Ag / AgCl is the reference electrode, platinum wire electrode is the auxiliary electrode, and the thyroglobulin is detected on the electrochemical workstation, and the electrochemical workstation and the chemiluminescence detector are combined Connect and set the high voltage of the photomultiplier tube to 700 V, the cyclic voltammetry scanning potential range is 0 ~ 1.6 V, and the scanning rate is 0.15 V / s; (2) In PBS with pH 8.0, by electrochemiluminescence method, the 1×10 -1 μg / mL~1×10 -7 Measure the electrochemiluminescence signal intensity generated by a series of thyroglobulin standard solutions with different concentrations on the modified electrod...

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Abstract

The invention discloses a photo-thermal release signal enhanced type thyroglobulin electrochemiluminescence immunosensor. According to a construction method of the sensor, a silicon-based titanium dioxide magnetic bead is used as a substrate to fix a thyroglobulin antibody, mesoporous silica marked thyroglobulin adsorbing bipyridyl ruthenium is used as a signal probe, and a competitive immunosensor is constructed through specific recognition between an antigen and the antibody. An electrode for modifying the antibody is placed into a mixed solution containing different concentrations of free antigens and a certain amount of signal probes, so that the free target antigen and the signal probe compete to bind the antibody, and a certain luminous intensity can be generated by combining the electrode of the signal probe. Under the irradiation of an 808 nm infrared laser, a sensing interface is rapidly heated, the dipyridyl ruthenium adsorbed by the mesoporous silica is released, and a luminous signal is remarkably enhanced along with the temperature rise, so that signal amplification is realized. As the concentration of the free target is increased, the number of the signal probes combined with the antibody is reduced, the luminous intensity is also reduced, and an electrochemiluminescence analysis method for the thyroglobulin can be established on the basis of the phenomenon.

Description

technical field [0001] The invention belongs to the technical field of novel functional materials and biosensing detection, and in particular relates to a method for detecting thyroglobulin based on a photothermal release signal enhanced electrochemiluminescence immunosensor. Background technique [0002] Electrochemiluminescence immunosensor, which combines electrochemiluminescence (ECL) with immunoassay, can be used for clinical diagnosis, and has the advantages of high sensitivity, strong specificity, good selectivity, and easy operation. The introduction of co-reactants into the ECL system can significantly enhance the intensity of its luminescent signal and effectively improve the sensitivity of the immunosensor. The present invention first proposes tetraethylenepentamine as a new co-reaction reagent of bipyridyl ruthenium, combined with the photothermal release effect of mesoporous silica, which significantly enhances the ECL signal and realizes highly sensitive detect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N27/48G01N27/30G01N27/327
CPCG01N33/68G01N27/3275G01N27/308G01N27/305G01N27/48
Inventor 戴宏任卉竹赵丹丹张书培高利红
Owner FUJIAN NORMAL UNIV
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