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A kind of preparation method of quantum dot-antibody immune complex

A technology of immune complexes and quantum dots, applied in measurement devices, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of increased preparation cost, different methods, low coupling rate, etc., and is conducive to large-scale production, Effect of Surface Potential Reduction and Molecular Weight Increase

Active Publication Date: 2021-06-11
RES INST OF XIAN JIAOTONG UNIV & SUZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the prior art, there are different methods for preparing quantum dot-antibody immune complexes, resulting in large differences in coupling efficiency and increased preparation costs
The current mainstream preparation method is: react with an activator first, and then combine with a specific antibody to prepare a quantum dot-antibody immune complex; however, according to the current specific preparation method, the quantum dot-antibody immune complex is even The coupling rate is generally low, which wastes raw materials and is not conducive to large-scale production, which limits its application in bioluminescence labeling

Method used

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  • A kind of preparation method of quantum dot-antibody immune complex
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  • A kind of preparation method of quantum dot-antibody immune complex

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Embodiment 1

[0035]Dissolve 5 μL of quantum dots (emission peak at 625nm±5nm, concentration 8 μM, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd.) in 500 μL, pH=5.5, 0.01M boric acid-borax buffer solution, vortex to mix Uniform 30s. Now prepare 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and 0.01M N-hydroxysulfosuccinimide (suLfo-NHS), solvent It is pH=5.5, 0.01M boric acid-borax buffer solution. Add 8 μL, 0.01M EDC to the quantum dot solution, vortex and mix, and add 40 μL, 0.01M suLfo-NHS after 5 minutes. Sonicate at 1±1°C for 30 min. After the activation is completed, take out the activation reaction solution, place it in an ultrafiltration tube (100kd), and centrifuge in a low-temperature centrifuge at a speed of 3500rpm for 5min. After completion, add 1ml, pH=8.5, 0.01M boric acid-borax to the reaction solution buffer solution, repeated centrifugation once, and removed the reaction solution. Add 7.2 μL of mouse CRP monoclonal ...

Embodiment 2

[0041] Dissolve 5 μL of quantum dots (emission peak at 625nm±5nm, concentration 8 μM, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd.) in 500 μL, pH=5.5, 0.01M boric acid-borax buffer solution, vortex to mix Uniform 30s. Now prepare 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and 0.01M N-hydroxysulfosuccinimide (suLfo-NHS), solvent It is pH=5.5, 0.01M boric acid-borax buffer solution. Add 16 μL, 0.01M EDC to the quantum dot solution, vortex and mix, add 40 μL, 0.01M suLfo-NHS after 5 min, and vortex mix. Sonicate at 1±0.5°C for 30min. After the activation is completed, take out the activation reaction solution, place it in an ultrafiltration tube (100kd), and centrifuge in a low-temperature centrifuge at a speed of 3500rpm for 5min. After completion, add 1ml, pH=8.5, 0.01M boric acid-borax to the reaction solution buffer solution, repeated centrifugation once, and removed the reaction solution. Add 7.2 μL of mouse CRP ...

Embodiment 3

[0046] Dissolve 5 μL of quantum dots (emission peak at 625nm±5nm, concentration 8 μM, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd.) in 500 μL, pH=5.5, 0.01M boric acid-borax buffer solution, vortex to mix Uniform 30s. Now prepare 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and 0.01M N-hydroxysulfosuccinimide (suLfo-NHS), solvent It is pH=5.5, 0.01M boric acid-borax buffer solution. Add 16 μL, 0.01M EDC to the quantum dot solution, vortex and mix, add 40 μL, 0.01M suLfo-NHS after 5 min, and vortex mix. Sonicate at 1±1°C for 30 min. After the activation is completed, take out the activation reaction solution, place it in an ultrafiltration tube (100kd), and centrifuge in a low-temperature centrifuge at a speed of 3500rpm for 5min. After completion, add 1ml, pH=8.5, 0.01M boric acid-borax to the reaction solution buffer solution, repeated centrifugation once, and removed the reaction solution. Add 5.2 μL of mouse CRP m...

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Abstract

The invention discloses a preparation method of a quantum dot-antibody immune complex. The preparation method comprises the following steps: 1) buffering the carboxyl group-modified quantum dots and an activator at a pH of 5-6 at 0-10° C. Reaction in liquid to generate activated quantum dots; wherein, the activator comprises 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, N-hydroxysuccinimide or its thiol 2) The activated quantum dots prepared in step 1) are reacted with the antibody at 0-10° C. in a buffer with a pH value of 6-9 to prepare a quantum dot-antibody immune complex; the present invention can realize the quantum dots – Antibody immune complexes have the characteristics of strong fluorescence intensity and biological activity, and can achieve a coupling rate of about 90%, which greatly improves the utilization of raw materials and is conducive to large-scale applications.

Description

technical field [0001] The invention belongs to the field of biomarkers, and in particular relates to a preparation method of a quantum dot-antibody immune complex. Background technique [0002] Quantum dots (QDs) are semiconductor nanoparticles mainly composed of group IIB~VIA elements (such as CdSe, CdTe, CdS, ZnSe, etc.) or group IIIA~VA elements (such as InP, InAs, etc.). Since the synthesis was discovered in the 1980s, due to its special fluorescence characteristics, such as: 1) the size controls the emission spectrum, the color is adjustable, and the multicolor labeling of the same quantum dot can be realized; 2) the excitation spectrum is wide, and the emission spectrum is narrow and symmetrical ; 3) Larger Stokes shift; 4) High photochemical stability and resistance to photobleaching; 5) High fluorescence efficiency; 6) Good biocompatibility; 7) Long fluorescence lifetime, etc., so it can be extended to be used in biological mark. [0003] At present, quantum dots ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/536G01N21/64
CPCG01N21/6428G01N33/533G01N33/536G01N2021/6432
Inventor 李嫚莉李超胡延祯
Owner RES INST OF XIAN JIAOTONG UNIV & SUZHOU
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