A high-sensitivity immunoassay for C-reactive protein based on a two-color quantum dot ratiometric fluorescent probe

A technology of ratiometric fluorescent probes and immunoassays, applied in fluorescence/phosphorescence, measurement devices, biological testing, etc., can solve the problems of randomness in the ratio of fluorescence intensity of microspheres, low loading efficiency, fluorescence quenching, etc., to avoid Effect of Luminous Efficiency Decrease

Active Publication Date: 2022-05-17
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above method requires some modification of the quantum dots, so the ligands on the surface of the quantum dots may fall off during the assembly process, resulting in fluorescence quenching, while the loading efficiency is low, and the fluorescence intensity ratio of the final microspheres is random.

Method used

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  • A high-sensitivity immunoassay for C-reactive protein based on a two-color quantum dot ratiometric fluorescent probe
  • A high-sensitivity immunoassay for C-reactive protein based on a two-color quantum dot ratiometric fluorescent probe
  • A high-sensitivity immunoassay for C-reactive protein based on a two-color quantum dot ratiometric fluorescent probe

Examples

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Embodiment 1

[0039] Add 0.068 g of triethanolamine (TEA) to 25 mL of ultrapure water, and gently magnetically stir the reaction in an oil bath at 80 °C for 30 min. Add 0.38 g of cetyltrimethylammonium bromide (CTAB) and 0.168 g Add sodium salicylate (NaSal) to the above solution, continue stirring for 1 h, then add 4 mL tetraethylorthosilicate (TEOS) to the above water-CTAB-NaSal-TEA solution, continue stirring for 3 h Afterwards, the product was washed three times with absolute ethanol and dispersed in ethanol. After the product from the previous step was ultrasonicated into a homogeneous phase, it was centrifuged and filtered, and the obtained product was added to a mixture of 50 mL of hydrochloric acid and 50 mL of methanol, and magnetically stirred in a water bath at 60°C for 6 h to react twice, and then the product was washed with absolute ethanol Three times, dendritic silica microspheres with an average diameter of 300nm and an average pore diameter of 25nm were obtained (the averag...

Embodiment 2

[0042] Add 0.068 g of triethanolamine (TEA) into 25 mL of ultrapure water, and gently magnetically stir the reaction in an oil bath at 80 °C for 30 min. Add 0.38 g of cetyltrimethylammonium bromide (CTAB) and 0.252 g Add sodium salicylate (NaSal) to the above solution, continue stirring for 1 h, then add 4 mL tetraethylorthosilicate (TEOS) to the above water-CTAB-NaSal-TEA solution, continue stirring for 3 h Afterwards, the product was washed three times with absolute ethanol and dispersed in ethanol. After the product from the previous step was ultrasonicated into a homogeneous phase, it was centrifuged and filtered, and the obtained product was added to a mixture of 50 mL of hydrochloric acid and 50 mL of methanol, and magnetically stirred in a water bath at 60 °C for 6 h to react twice, and then the product was washed with absolute ethanol Three times to obtain dendritic silica microspheres with an average diameter of 300 nm and an average pore diameter of 30 nm. The resul...

Embodiment 3

[0044] Add 0.068 g of triethanolamine (TEA) into 25 mL of ultrapure water, and gently stir the reaction with magnetic force in an oil bath at 80 °C for 30 min. Add 0.38 g of cetyltrimethylammonium bromide (CTAB) and 0.286 Add sodium salicylate (NaSal) to the above solution, continue stirring for 1 h, then add 4 mL tetraethylorthosilicate (TEOS) to the above water-CTAB-NaSal-TEA solution, continue stirring for 3 h Afterwards, the product was washed three times with absolute ethanol and dispersed in ethanol. After the product from the previous step was ultrasonicated into a homogeneous phase, it was centrifuged and filtered, and the obtained product was added to a mixture of 50 mL of hydrochloric acid and 50 mL of methanol, and magnetically stirred in a water bath at 60°C for 6 h to react twice, and then the product was washed with absolute ethanol Three times, dendritic silica microspheres with an average diameter of 300 nm and an average pore diameter of 32 nm were obtained. ...

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Abstract

The invention discloses a high-sensitivity immunodetection method for C-reactive protein based on a two-color quantum dot ratio fluorescent probe, which couples silver nanoparticles AgNPs to a CRP-labeled antibody, and then performs double-antibody sandwiching of C-reactive protein with a capture antibody to obtain Double-antibody sandwich model; further utilization of cyanide CN ‑ Dissolving the AgNPs captured by the double-antibody sandwich model allows the AgNPs to release Ag + The signal is amplified, and then the two-color quantum dot ratio fluorescent probe is added to measure the fluorescence spectrum to realize the quantitative analysis of C-reactive protein; the two-color quantum dot ratio fluorescent probe is based on red quantum dots and embedded in The two-color fluorescent microspheres are formed by thiol dendritic silica balls, and green quantum dots are assembled on the outer layer. The method of the present invention utilizes CN ‑ Corrosion of AgNPs to make them into Ag + , based on the ion exchange between silver ions and quantum dot ratio fluorescent probes, the concentration of C-reactive protein can be detected qualitatively and quantitatively through the change of fluorescent signal and solution color.

Description

technical field [0001] The invention relates to a high-sensitivity immunodetection method for C-reactive protein based on a two-color quantum dot ratio fluorescent probe. Background technique [0002] Ratiometric fluorescent probe is a new type of fluorescent sensor that has received extensive attention in recent years. Ratiometric fluorescent probe usually refers to a fluorescent sensor with dual emission characteristics constructed using two fluorescent materials with different emission wavelengths. According to the ratio of the two emission peaks Changes, establish the relationship with the content of the target substance, and conduct quantitative and qualitative analysis of the target substance. For the detection of target objects using a single fluorescent channel, only the signal at the corresponding emission wavelength is analyzed, because some unavoidable factors that have nothing to do with the target object will affect the results, such as: light scattering of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/533G01N21/64
CPCG01N33/68G01N33/54346G01N33/533G01N21/6428G01N2021/6432Y02A50/30
Inventor 黄亮吴枫汪晶
Owner ZHEJIANG UNIV OF TECH
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