Preparation method for quantum dot-cardiac troponin I antibody immune complex, and preparation method for test strip

A technique for cardiac troponin and immune complexes, which is applied in the field of biomarkers, can solve the problems of unable to obtain a standard curve of myocardial markers, unable to realize the detection of myocardial markers, etc., and achieves good coupling effect, simple preparation method, and standard curve. Good results

Inactive Publication Date: 2019-08-02
RES INST OF XIAN JIAOTONG UNIV & SUZHOU
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] CN104280545A discloses a multi-indicator quantum dot marking test strip and method for synchronously quantifying myocardial markers. However, the inventor found through research that the standard curve of the corresponding myocardial markers cannot be obtained only by using the content disclosed in this patent, so that it cannot Realize the detection of corresponding myocardial markers

Method used

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  • Preparation method for quantum dot-cardiac troponin I antibody immune complex, and preparation method for test strip
  • Preparation method for quantum dot-cardiac troponin I antibody immune complex, and preparation method for test strip
  • Preparation method for quantum dot-cardiac troponin I antibody immune complex, and preparation method for test strip

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Preparation of a quantum dot-cardiac troponin I antibody immune complex:

[0046] Dissolve 5ul of quantum dots (average particle size of 18 nm, emission peak of 625nm±5nm, concentration of 8uM, purchased from Wuhan Jiayuan) in 500ul, pH 5.5, 0.01M borate buffer, vortex and mix for 30s . Now prepare 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 0.01M N-hydroxysulfosuccinimide, the solvent is pH 5.5, 0.01M boric acid salt buffer. Add 0.8 ul, 0.01M 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride to the quantum dot solution, vortex and mix, add 40ul, 0.01M N- Hydroxysulfosuccinimide, vortexed to mix. Sonicate in ice bath 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 3500 rpm for 7 minutes. After completion, add 1ml of borate buffer, pH 8.5, 0.01M, to the reaction solution solution, rep...

Embodiment 2

[0056] Basically the same as in Example 1, the difference is that: when preparing the quantum dot-cardiac troponin I antibody immune complex, borate buffers with different pH were used to adjust the pH of the reaction solution after activation, and the agarose gel Electropherogram see figure 2 , wherein, 1 is the band of the quantum dot-cardiac troponin I antibody immune complex prepared by the borate buffer solution of pH 5.5, and 2 is the quantum dot- The band of the immune complex of cardiac troponin I antibody, 3 is the band of the quantum dot-cardiac troponin I antibody immune complex prepared in borate buffer solution at pH 7.5, and 4 is the boron at pH 8.5 The bands of the quantum dot-cardiac troponin I antibody immune complex prepared in salt buffer solution, 5 are the bands of the quantum dot-cardiac troponin I antibody immune complex prepared in the borate buffer solution of pH9.5 Band, 6 is the band of the control.

[0057] Analysis Due to the different isoelectr...

Embodiment 3

[0060] Basically the same as Example 1, the difference is: when preparing the quantum dot-cardiac troponin I antibody immune complex, respectively adding different molar ratios of quantum dots (QDs) and 1-ethyl-3-(3- Dimethylaminopropyl) carbodiimide hydrochloride (EDC), agarose gel electrophoresis image 3 , wherein, 1 is the band of contrast; 2 is the band that the molar ratio of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride is 1:200, 3 is the strip with the molar ratio of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride being 1:1000, and 4 is the band of quantum dots and 1-ethyl- 3-(3-Dimethylaminopropyl)carbodiimide hydrochloride is a band with a molar ratio of 1:5000, 5 is quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) The molar ratio of carbodiimide hydrochloride is 1:25000, and 6 is the band of quantum dots and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. Bands at a molar ratio of 1:5000...

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Abstract

The invention relates to a preparation method for a quantum dot-cardiac troponin I antibody immune complex. A quantum dot and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-Hydroxysuccinimide orthe sulfo-product thereof react so as to control the charge molar ratio of the quantum dot, the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, the N-Hydroxysuccinimide or the sulfo-product thereof tobe 1:200-1000:5000-10000; regulating the pH (Potential of Hydrogen) of reaction liquid to be 8-9; and adding the cardiac troponin I antibody into the reaction liquid, and controlling the charge molarratio of the quantum dot to the cardiac troponin I antibody to be 1:42-54. The preparation method has the advantages of good coupling effect, simple preparation method and low cost. The prepared teststrip has a good standard curve, a relevant coefficient is greater than 0.99, detection sensitivity is high, and the accurate quantitative detection of the cardiac troponin I antibody can be realized.

Description

technical field [0001] The invention belongs to the field of biomarkers, and in particular relates to a preparation method of a quantum dot-cardiac troponin I antibody immune complex and a preparation method of a test strip. 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.), three The dimensions are all below 100nm, and the appearance is just like a dot. Quantum dots synthesized in aqueous phase have good biocompatibility and are easy to introduce functional groups. Compared with traditional organic conventional fluorescent materials, quantum dots have special fluorescent characteristics: 1) The emission spectrum can be controlled by changing the size and composition, and the color is adjustable, which can realize multi-color labeling of the same quantum dot; 2) Organic dye fluorescence The molecular excitation l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532
CPCG01N33/532
Inventor 胡延祯李超
Owner RES INST OF XIAN JIAOTONG UNIV & SUZHOU
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