Kit for early diagnosis of Alzheimer's disease and application thereof
A technology for early diagnosis of Alzheimer's disease, applied in disease diagnosis, biological testing, material inspection products, etc., can solve the problems of lack of accurate prediction ability of Alzheimer's disease, long detection time, complicated operation, etc., and achieve easy The effect of operation, wide detection range and simple operation
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[0030] In one embodiment of the present invention, a kind of preparation method of above-mentioned kit is provided, and described preparation method comprises:
[0031] The preparation method of reagent A is as follows: T-Tau, p-Tau-181, p-Tau-217 and p-Tau-231 monoclonal antibodies are mixed with biotin at a molar ratio of 1:10-1:40 Mix well at room temperature and react for 1-4 h, then dialyze the reactant in PBS buffer to obtain biotin-labeled monoclonal antibody; add biotin-labeled monoclonal antibody to the buffer of streptavidin magnetic microspheres In the system, the coupling ratio of biotinylated protein to streptavidin magnetic microspheres is 10-30μg:1mg.
[0032] The preparation method of reagent B is as follows: mix the monoclonal antibody of Tau protein N 1-17 site and acridinium ester according to the molar ratio of 1:10-1:40, react at room temperature for 2-4 h, and then add lysine The coupling reaction was terminated by acid, and the reaction product was obta...
Embodiment 1
[0041] In this embodiment, a kit for early diagnosis of Alzheimer's disease is provided, which includes: reagent A1 (T-Tau magnetic microsphere working solution), A2 (p-Tau-181 magnetic microsphere Ball working solution), A3 (p-Tau-231 magnetic microsphere working solution), A4 (p-Tau-217 four kinds of magnetic microsphere working solution), reagent B1 (monoclonal Antibody-prepared acridinium ester working solution), calibrator (universal composite calibrator, used to establish a standard curve).
[0042] Step 1: Buffer configuration
[0043] 1. The preparation method of the buffer system of magnetic microspheres
[0044] In the 0.15 M PBS buffer system, adjust the pH to 7.3, add 3% BSA, 2% trehalose, 0.05% magnetic microsphere dispersant, 0.25% Tween 20, 0.05% Proclin 300, mix well until completely dissolved, Add an appropriate amount of purified water to quantify, stir and mix for 1 h, and adjust the pH to 7.3. Filter it with a filter device and store it at 4°C for future...
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