A detection reagent and its preparation method and application
A detection reagent and a reagent technology, applied in the field of detection reagents and their preparation, can solve problems such as poor detection sensitivity of chemiluminescence method, and achieve the effects of improving the value and sensitivity of reaction signals, enhancing sensitivity, and enhancing sensitivity
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[0053] On the other hand, the embodiment of the present invention also provides a method for preparing the detection reagent described above, including:
[0054] A cross-linking reagent and an enzyme-labeled antibody are provided, and the cross-linking reagent and the enzyme-labeled antibody are reacted in a solution to prepare an enzyme-labeled antibody polymer.
[0055] Specifically, the cross-linking reagent refers to a type of reagent that bonds and cross-links two or more molecules to each other to form a relatively stable multi-molecular polymer. Any cross-linking reagent that can link multiple enzyme-labeled antibodies to each other to form a multimolecular polymer and provide carbon bridges for the linkage between enzyme-labeled antibodies can be used in the embodiments of the present invention.
[0056] Preferably, the crosslinking agent is a homobifunctional crosslinking agent. Homobifunctional cross-linking agent is a kind of reagent that has the same active group ...
Embodiment 1
[0073] This embodiment provides a chemiluminescent detection kit, including: magnetic separation reagents, enzyme-labeled reagents, chemiluminescent substrate solutions and calibration products, the preparation of which includes:
[0074] 1. Preparation of enzyme-labeled reagents
[0075] (1) Select troponin T monoclonal antibody as the detection antibody, add 12mg / mL 2-iminothiolane hydrochloride solution according to 1 / 100 of the volume of the antibody, mix well, and place it at room temperature for 30min to activate treatment to obtain activated antibodies;
[0076] Select alkaline phosphatase as the labeled enzyme, add 7mg / mL Sulfo-SMCC solution according to 1 / 10 of the volume of alkaline phosphatase, mix well, place at room temperature for 25min, and perform activation treatment to obtain activated alkaline phosphatase solution;
[0077] The activated alkaline phosphatase and the activated troponin T monoclonal antibody were mixed and reacted at 4° C. for 15 hours to prep...
Embodiment 2
[0085] This embodiment provides a chemiluminescent detection kit, including: magnetic separation reagents, enzyme-labeled reagents, chemiluminescent substrate solutions and calibration products, the preparation of which includes:
[0086] 1. Preparation of enzyme-labeled reagents
[0087] (1) Select troponin T monoclonal antibody as detection antibody, select horseradish base peroxidase (HRP) as labeling enzyme;
[0088] Centrifuge the HRP and add fresh 0.3M NaIO 4 The solution was stirred at room temperature for 50 minutes in the dark to obtain the hydroformylated HRP; ultrafiltration was performed using 0.05M, pH 4.4 sodium acetate buffer solution, and the ultrafiltration conditions were set at 4°C, 9000rmp, 20min, ultrafiltration 3 times, and the purified Add 1M, pH 9.5 carbonate buffer, adjust the pH to 9.0-9.5, then add troponin T monoclonal antibody, shake at room temperature for 7 hours in the dark; add 0.1ml of newly prepared 9mg / mL of NaBH 4 solution, shake for 2h...
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