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Quantum dot-antibody fluorescent probe, preparation method, probe and test paper strip

A fluorescent probe and quantum dot technology, applied in the detection field, can solve the problems of high toxicity, low luminous efficiency, poor uniformity and dispersion, etc., and achieve the effects of high detection sensitivity, high fluorescence recovery rate, and high detection accuracy

Inactive Publication Date: 2017-03-22
JIANGSU LIANGDIAN TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The quantum dots synthesized in organic systems have almost perfect crystal structure, good monodispersity, strong photostability and high fluorescence yield, but tributylphosphine, trioctyl Organic phosphine compounds such as phosphine are highly toxic, flammable, unstable, explosive, expensive, and have high requirements for the operating environment, which limits the large-scale production and application of quantum dot products
In addition, in the process of preparing quantum dot fluorescent probes, the coupling products of quantum dots and corresponding antibody molecules have poor uniformity and dispersion, are prone to agglomeration, low luminous efficiency, and the half-peak width of the emission spectrum is too broad, etc. Problems that affect the detection performance of related products

Method used

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  • Quantum dot-antibody fluorescent probe, preparation method, probe and test paper strip
  • Quantum dot-antibody fluorescent probe, preparation method, probe and test paper strip
  • Quantum dot-antibody fluorescent probe, preparation method, probe and test paper strip

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preparation example Construction

[0021] One aspect of the present invention provides a method for preparing a quantum dot-antibody fluorescent probe, which includes the following steps:

[0022] (1) Add quantum dots to borate buffer solution to prepare quantum dot solution;

[0023] (2) The activator is added to the quantum dot solution to activate the quantum dots, the activator is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or its hydrochloride and N -Hydroxysuccinimide or its thiol, the molar ratio of the activator to the quantum dot is 5-9:1, wherein the 1-(3-dimethylaminopropyl)-3-ethylcarbodi The molar ratio of imine or its hydrochloride to N-hydroxysuccinimide or its sulfide is 2~3:4~5;

[0024] (3) The antibody is added to the activated quantum dot solution for coupling, and the molar ratio of the antibody to the quantum dot is 4-2:1 to obtain a quantum dot-antibody fluorescent probe.

[0025] The quantum dot-antibody fluorescent probe prepared by the method has high fluorescence recovery rate, and the rec...

Embodiment 1

[0070] Weigh 20mg CdSe / ZnS QDs and dissolve in 4ml chloroform solution, weigh 20mg maleic anhydride-octadecene copolymer and dissolve in 2ml chloroform, respectively ultrasonically dissolve them, mix the two solutions with ultrasonic for 5~10min , Rotary evaporation to remove the chloroform solvent, the quantum dots are in a dry state, add 4ml of aqueous solution, ultrasonic for 30min to dissolve, the solution is clear and bright.

Embodiment 2

[0072] Add 150ul of carboxyl water-soluble quantum dots prepared in Example 1 (concentration is 1mg / mL), add 250ul pH 7.4, 20mM borate buffer, vortex mixer to mix; add 50mg / ml EDC 50ul, vortex Mix with a rotary mixer, add 50ul of 100mg / ml NHS, vortex to mix; ice-bath ultrasonic activation for 5min; add antibody 50.3ug, vortex mixer to mix; place on a rotary incubator at 37°C for 2h. Add 3ul ethanolamine (concentration is 100%) and 50ul 20% BSA, mix well; place it on a rotating incubator at 37°C for 30 minutes, centrifuge at a speed of 22000 rpm, centrifuge for 15 minutes, discard the supernatant, and repeat twice; combine the precipitate to dissolve 150ul pH 8.0, 10mM borate buffer solution is quantum dot-antibody fluorescent probe.

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Abstract

The invention provides a quantum dot-antibody fluorescent probe preparation method, a quantum dot-antibody fluorescent probe, and an immunochromatography test paper strip and a preparation method thereof. The quantum dot-antibody fluorescent probe preparation method comprises: (1) adding quantum dots to a borate buffer solution to prepare a quantum dot solution; (2) adding an activator to the quantum dot solution, and activating, wherein the activator is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or a hydrochloride thereof and N-hydroxysuccinimide or a sulfide thereof, a molar ratio of the activator to the quantum dots is 5-9:1, and a molar ratio of the 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or the hydrochloride thereof to the N-hydroxysuccinimide or the sulfide thereof is 2-3:4-5; and 3) adding antibody to the activated quantum dot solution, and coupling to obtain the quantum dot-antibody fluorescent probe, wherein a molar ratio of the antibody to the quantum dots is 4-2:1.

Description

Technical field [0001] The invention relates to the field of detection, in particular to a method for preparing a quantum dot-antibody fluorescent probe, a quantum dot-antibody fluorescent probe, and an immunochromatographic test strip containing the probe. Background technique [0002] Quantum dots (QDs), also known as semiconductor nanocrystals, are usually composed of II-VI and III-V elements, with adjustable size, wide excitation spectrum, narrow emission spectrum, large Stoke shift, etc. Features. The molar absorption coefficient and fluorescence quantum yield of quantum dots are relatively high, which can generate strong fluorescence signals and improve detection sensitivity. At the same time, quantum dots have good photochemical stability and good photobleaching resistance, which improves the stability of detection results. In summary, quantum dots are an ideal fluorescent probe material, and have unique advantages and broad application prospects in the detection field b...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/558G01N33/533
CPCG01N33/582G01N33/533G01N33/558G01N33/588
Inventor 陈婷李林松柴向东李金洁
Owner JIANGSU LIANGDIAN TECH CO LTD
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