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Fluorescence immune probe as well as preparation method and application thereof

A fluorescent immunity and probe technology, applied in the field of fluorescent immunity probes and their preparation, can solve the problems of long preparation period and low sensitivity of quantum dot fluorescent immunity probes, shorten the preparation period, improve the reaction efficiency, improve the sensitivity and The effect of stability

Pending Publication Date: 2022-07-05
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies and actual needs of the prior art, the present invention provides a fluorescent immunoprobe and its preparation method and application. The method can rapidly prepare a highly sensitive quantum dot fluorescent immunoprobe, effectively solving the problem of current quantum dot fluorescent immunoprobes. Problems such as long needle preparation cycle and low sensitivity

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  • Fluorescence immune probe as well as preparation method and application thereof
  • Fluorescence immune probe as well as preparation method and application thereof
  • Fluorescence immune probe as well as preparation method and application thereof

Examples

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Embodiment 1

[0067] This embodiment provides a quantum dot fluorescent immunoprobe, and the preparation method for preparing the quantum dot fluorescent immunoprobe includes the following steps:

[0068] (1) Add coupling solution, 50 μL 54 mg / mLEDC and 50 μL 75.6 mg / mL NHS to 150 μL of 5 mg / mL quantum dot solution, sonicate for 5 min, and then centrifuge to remove the supernatant to retain the precipitate;

[0069] (2) The coupling solution was added to the precipitation in step (1), then 21 μL of 10 mg / mL streptavidin was added, and the coupling was shaken at 200 rpm for 0.7 h;

[0070] (3) Add blocking solution to the solution obtained in step (2), and react at 25°C for 0.5h

[0071] (4) centrifuging the solution obtained in step (3) to remove the supernatant and retaining the precipitate, and redissolving with the preservation solution to obtain a quantum dot-streptavidin conjugate;

[0072] (5) Biotin was added to the buffer containing cortisol antibody, the molar ratio of biotin and ...

Embodiment 2

[0080] This embodiment provides a quantum dot fluorescent immunoprobe, and the preparation method of the quantum dot fluorescent immunoprobe is compared with Embodiment 1, the only difference is that the quantum dot-streptavidin couple in step (8) The molar ratio of conjugate and biotinylated antibody was 1:1.

Embodiment 3

[0082] This embodiment provides a quantum dot fluorescent immunoprobe, and the preparation method of the quantum dot fluorescent immunoprobe is compared with Embodiment 1, the only difference is that the quantum dot-streptavidin couple in step (8) The molar ratio of conjugate and biotinylated antibody was 1:10.

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Abstract

The invention discloses a fluorescence immunoassay probe as well as a preparation method and application thereof. The method for preparing the quantum dot fluorescence immunoassay probe comprises the following steps: mixing a quantum dot-streptavidin conjugate, a biotinylated antibody and a borate buffer solution, and carrying out oscillatory reaction to obtain the quantum dot fluorescence immunoassay probe. According to the preparation method disclosed by the invention, the quantum dot fluorescence immunoassay probe is prepared through a reaction system of streptavidin and biotin in combination with a borate buffer solution and a high-speed oscillation reaction, so that the high-sensitivity quantum dot fluorescence immunoassay probe can be rapidly prepared; the problems that an existing quantum dot fluorescence immunoassay probe is long in preparation period, low in sensitivity and the like are effectively solved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a fluorescent immune probe and a preparation method and application thereof. Background technique [0002] Immunolabeling technology refers to the use of chromogenic and luminescent substances as indicators to label antigens or antibodies through covalent or non-covalent bonds to carry out antigen-antibody reactions. Qualitative and quantitative determination of the experimental results. A major breakthrough was made in trace analytical chemistry, combining radioisotope tracing with immunoreaction to establish radioimmunoassay. Since then, immunoassay technologies have been continuously developed and made breakthroughs, such as enzyme-linked immunosorbent assay, colloidal gold immunochromatography, fluorescence immunoassay, and chemiluminescence immunoassay. [0003] During the synthesis of fluorescent immunoprobes, it is usually necessary to couple proteins (antigens or antibodies, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/58G01N33/533
CPCG01N33/588G01N33/533
Inventor 柴语鹃李启航胡晓祥郑钧元曾雨萍
Owner SHENZHEN UNIV