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Two-photon fluorescence labeling probe, and synthesis method thereof and application of two-photon fluorescence labeling probe in novel coronavirus diagnosis

A two-photon fluorescence and labeling probe technology, which is applied in the direction of fluorescence/phosphorescence, chemical instruments and methods, and luminescent materials, to achieve the effects of improving fluorescence quantum yield, simple structure, and low production cost

Active Publication Date: 2020-08-07
ZUNYI NORMAL COLLEGE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the research process of the two-photon fluorescent probe, the researchers of the present invention have focused on the detection of intracellular mercury ions, zinc ions, silver ions, sodium ions, etc., the activity detection of intracellular sugar chain antigens and temperature sensing detection, etc. All aspects have been studied, but the technical solution for the biopsy of the antigen of the new coronavirus 2019-nCoV based on the combination of the naphthalene structure parent and the hydroxysuccinimide ester has not been reported

Method used

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  • Two-photon fluorescence labeling probe, and synthesis method thereof and application of two-photon fluorescence labeling probe in novel coronavirus diagnosis
  • Two-photon fluorescence labeling probe, and synthesis method thereof and application of two-photon fluorescence labeling probe in novel coronavirus diagnosis
  • Two-photon fluorescence labeling probe, and synthesis method thereof and application of two-photon fluorescence labeling probe in novel coronavirus diagnosis

Examples

Experimental program
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Effect test

Embodiment 1

[0064] The synthetic method of two-photon fluorescent labeling probe LP, comprises the steps:

[0065] (1) Synthesis of 6-acetyl-2-naphthol (referred to as compound 2) (references are carried out): add 20 grams of 6-methoxyl-2-acetylnaphthalene (abbreviated as compound 1) and 200 ml of glacial acetic acid, stirred, then added 86 g of hydrobromic acid, and stirred and refluxed at 100°C for 12 hours, the black reaction solution was desolvated under reduced pressure at 60°C, neutralized by adding 10% sodium bicarbonate solution, and washed with ethyl acetate Extraction, organic phase with anhydrous MgSO 4 Drying, filtration, solvent removal under reduced pressure, column separation, 8.03g of 43.17mmol yellow powder is compound 2, yield 43%; wherein, the eluent used for column separation is V (petroleum ether): V (ethyl acetate )=2:1; the structure of the compound 2 is consistent with that reported in the literature (H.M.Kim, C.Jung, B.R.Kim, et al., Angew.Chem. Int.Ed., 2007,46:...

Embodiment 2

[0078] Example 2 Two-photon fluorescent labeling probe labeling N protein antigen Ag

[0079] Using Marsshall's method, the specific steps are as follows:

[0080] 1) Take 50 μL of 5 mg / mL N protein antigen solution, add 5 μL of carbonate buffer, 15 μL of 2 mg / mL LP-containing DMF solution in sequence, mix well, and react on a shaker at 200 rpm at 25°C for 2 hours; Carbonate buffer consists of Na 2 CO 3 8.6g,NaHCO 3 17.3g, prepared with 1000mL of distilled water, its pH is 9.3;

[0081]2) Purification by ultrafiltration: Take the liquid obtained in step 1) and add it to the ultrafiltration tube, add 0.02 mol / L, pH 7.2 phosphate buffer solution (PB) to 80% of the total volume of the ultrafiltration tube, and then in 4 ° C, 8000- Centrifuge at 10,000 rpm for 5-20 minutes, add PB repeatedly and centrifuge for 3-5 times to obtain the fluorescent antigen, namely LP-Ag precipitate.

Embodiment 3

[0082] Example 3 Immunochromatographic test strips for rapid detection of new coronavirus

[0083] The composition of the test strip: sample pad, binding pad, detection G line, detection M line, quality control C line, absorbent paper, PVC bottom plate; the sample pad contains blood filter membrane, and the binding pad is coated with the fluorescence obtained in Example 2. antigen.

[0084] The production method steps are as follows:

[0085] (1) Preparation of detection line and quality control line

[0086] Use Bio-Dot XYZ-3050 automatic film spraying machine to spray 1 mg / mL purified anti-human IgM secondary antibody, anti-human IgG secondary antibody, and goat anti-mouse IgG antibody on the nitrocellulose membrane, that is, the detection M line of the NC membrane, respectively. Detect the positions of the G line and the quality control C line at a scribing speed of 0.80 μL / cm, dry at 35°C for 5 hours, and seal and store at 4°C for later use;

[0087] (2) The binding pad...

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Abstract

The invention belongs to the technical field of two-photon fluorescence labeling probes and application thereof. The invention particularly relates to a two-photon fluorescence labeling probe, and a synthesis method thereof and application of the two-photon fluorescence labeling probe in novel coronavirus diagnosis. The probe has the chemical name of 3-(N-methyl-N-(6-acetyl-2-naphthyl)amino)propionic acid-N-hydroxysuccinimide ester. The two-photon fluorescent probe has the advantages of high sensitivity and rapid and accurate detection; a fluorescent antigen obtained by labeling an N protein antigen of 2019-nCoV with the probe can be made into an immunochromatographic test strip or a detection kit, and can be used for rapid detection of novel coronavirus, and the test strip or the kit hasthe excellent characteristics of rapid detection, high detection consistency and high stability.

Description

technical field [0001] The invention belongs to the technical field of making two-photon fluorescent labeling probes, and in particular relates to two-photon fluorescent labeling probes for the express diagnosis of the new coronavirus, its synthesis method, and the application of preparing immunochromatographic test strips and kits. Background technique [0002] As a pathogen that causes human respiratory disease (2019-nCoV), the novel coronavirus (2019-nCoV) has been closely related to humans and animals for a long time. Coronavirus is the largest virus in the RNA virus family. Its genome is about 27-32kb in length and has an envelope. The Spike glycoprotein on the surface of the virus is shaped like a "crown (corona Latin)". Coronavirus. The new coronavirus belongs to the new coronavirus of the genus β. It has an envelope, and the particles are round or oval, often pleomorphic, with a diameter of 60-140nm. Its genetic characteristics are significantly different from those...

Claims

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Application Information

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IPC IPC(8): C07D207/46C09K11/06G01N21/64
CPCC07D207/46C09K11/06C09K2211/1011C09K2211/1029G01N21/6428G01N2021/6439
Inventor 黄池宝齐向武胡瑞峰
Owner ZUNYI NORMAL COLLEGE
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