Signal amplification quantum dot fluorescence immunoassay probe as well as preparation method and application thereof
A technique of fluorescence immunity and signal amplification, applied in measurement devices, material testing products, instruments, etc., can solve the problems of limited signal intensity improvement, complicated preparation method steps, etc., to achieve improved sensitivity and linear performance, sufficient fluorescent markers and antibodies , the effect of linear width
Pending Publication Date: 2021-11-19
ANHUI HUIBANG BIOLOGICAL ENG
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Problems solved by technology
However, this technology only achieves one amplification of the signal, and the improvement of signal strength is limited
[0006] The signal amplification fluorescent probe described in the patent with the publication number CN102565383A is [fluorescence-biotinylated antibody]-streptavidin-[fluorescence-polylysine-biotin], and its preparation method is relatively complicated. Requires biotin labeling of two substances
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Embodiment 1
[0033] Embodiment 1: The preparation of gastrin 17 (G-17) quantum dot immunofluorescence chromatography kit comprises the following steps:
[0034] (1) Preparation of G-17 fluorescent probe:
[0035] a. Add 1ml of morpholineethanesulfonic acid buffer (0.05M pH 5.5) to 10mg of quantum dot microspheres, then add N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl ) carbodiimide to a final concentration of 5 mM, and shake at room temperature in the dark for 30 minutes to obtain activated quantum dot microspheres;
[0036] b. Wash the activated quantum dot microspheres with 50mM pH 6.5 phosphate buffer, take 3mg of avidin and mix it well, and shake it at room temperature for 2h in the dark. After the reaction, add casein with a final concentration of 1% and 0.05 % Tween 20, to block the remaining active sites, shake at room temperature for 1 h in the dark, after completion, wash with 0.02M pH 7.4 PBS buffer containing 1% BSA, and resuspend to obtain 3mg / ml quantum dot-pro ...
example 1
[0073] Table 3 Example 1 Stability Experiment
[0074]
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The invention discloses a preparation method of a signal amplification quantum dot fluorescence immunoassay probe. The method comprises the following steps of activating carboxyl quantum dot fluorescent microspheres by using a condensing agent, and adding avidin and casein for reaction to form a quantum dot fluorescent microsphere-avidin compound; connecting the activated biotin with an antibody to obtain a biotinylated antibody; and mixing a biotinylated antibody and the quantum dot fluorescent microsphere-avidin compound for primary reaction, adding the quantum dot fluorescent microsphere-avidin compound, then adding the biotinylated antibody for secondary reaction, and circularly marking for multiple times to prepare the signal-amplified quantum dot fluorescence immunoassay probe. The invention also discloses the signal amplification quantum dot fluorescence immunoassay probe prepared by the preparation method and application of the signal amplification quantum dot fluorescence immunoassay probe in fluorescence immunoassay. The probe prepared by the method is sufficient in fluorescent marker and antibody and stable in structure, and has characteristics of strong capture capability, high sensitivity, wide linearity and the like when being used for immunodetection.
Description
technical field [0001] The invention relates to the technical field of rapid immune analysis, in particular to a signal amplification quantum dot fluorescent immunoprobe and its preparation method and application. Background technique [0002] Immunolabeling technology refers to the use of color-developing and luminescent substances as indicators to carry out antigen-antibody reactions by covalently or non-covalently labeling antigens or antibodies. Qualitative and quantitative determination of the experimental results. A major breakthrough occurred in trace analytical chemistry, combining radioisotope tracer and immune response to establish radioimmunoassay. Since then, immunoassay technology has continued to develop and break through, such as enzyme-linked immunosorbent assay, colloidal gold immunochromatography, fluorescent immunoassay, and chemiluminescence immunoassay. [0003] Due to different principles, the above-mentioned different marking techniques have their ow...
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IPC IPC(8): G01N33/533G01N33/543G01N33/58
CPCG01N33/533G01N33/54313G01N33/588
Inventor 陈竹吴才桂陈一凡陈永晨张海龙
Owner ANHUI HUIBANG BIOLOGICAL ENG