Preparation method of quantum dots-antibody immunocomplex

A technology of immune complexes and quantum dots, applied in measurement devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of increased preparation cost, different methods, low coupling rate, etc. The effect of reducing and increasing molecular weight

Active Publication Date: 2018-11-30
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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  • Preparation method of quantum dots-antibody immunocomplex
  • Preparation method of quantum dots-antibody immunocomplex
  • Preparation method of quantum dots-antibody immunocomplex

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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

[0042] 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

[0048] 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 a preparation method of quantum dots-antibody immunocomplex, including the steps of: 1) performing a reaction to carboxylic-modified quantum dots with an activator at 0-10 DEG C in a buffer solution with pH value being 5-6 to obtain activated quantum dots, wherein the activator includes 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide or a thio-substance thereof; 2) performing a reaction to the activated quantum dots with an antibody at 0-10 DEG C in a buffer solution with pH value being 6-9, thus preparing the quantum dots-antibody immunocomplex. The immunocomplex has strong fluorescence intensity, bioactivity and like features and can reach about 90% in coupling rate. The preparation method is greatly improved in raw material utilizationrate and is beneficial to large-scale application.

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 IIB-VIA elements (such as CdSe, CdTe, CdS, ZnSe, etc.) or IIIA-VIA elements (such as InP, InAs, etc.). Since the discovery of synthesis in the 1980s, due to its special fluorescence characteristics, such as: 1) size-controlled emission spectrum, adjustable color, can realize multicolor labeling of the same quantum dots; 2) wide excitation spectrum, narrow and symmetrical emission spectrum 3) Large 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 biological applications mark. [0003] At present, quantum dots are a new type of nanomaterials, and the preparation met...

Claims

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Application Information

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Patent Type & Authority Applications(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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