Kit or test strips for detecting salbutamol
A technology for salbutamol and kits, which is applied in the field of kits or test strips for detecting salbutamol, which can solve the problems of high cost, inapplicability to large-scale sample screening and detection, cumbersome operation steps, etc., and achieve low cost and convenient inspection methods , The effect of simple sample pretreatment process
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Embodiment 1
[0043] Embodiment 1, preparation albuterol hapten
[0044] One, the preparation of albuterol hapten
[0045] 1. Weigh 10 mg (42 μmol) of albuterol, place it in a 10 mL Erlenmeyer flask, add 5 mL of aqueous sodium bicarbonate solution with a concentration of 3% (mass ratio), stir fully to dissolve it completely, add 6.2 μL (42 μmol) of ethylene glycol di Glycidyl ether was stirred at room temperature for 4 h to obtain solution I.
[0046] 2. The mixed solution in step 1 is vacuum-dried, and the obtained solid is the product.
[0047] 2. Characterization of albuterol hapten
[0048] The product prepared in step 1 is subjected to elemental analysis, and the results are as follows:
[0049] C: 63.88; H: 8.58; N: 3.26; O: 24.57.
[0050]The results show that the product prepared in step one is the compound shown in formula (I).
[0051]
[0052] Formula (I).
Embodiment 2
[0053] Embodiment 2, preparation albuterol artificial antigen
[0054] 1. Preparation and characterization of albuterol immunogen
[0055] 1. Preparation of albuterol immunogen
[0056] Get 10mg of the compound shown in the formula (I) that embodiment 1 prepares, add bovine serum albumin solution (50mg bovine serum albumin is dissolved in 5mL PBS damping fluid) dropwise wherein, continue to stir and pack into dialysis bag after 8 hours, Dialyze in PBS buffer at 4°C for 72 hours (change the water 6 times in the middle), then centrifuge at 8000rmp for 30min, take the supernatant (salbutamol immunogen solution), aliquot into ampoule bottles, and store at -20°C. Salbutamol immunogen is referred to as SAL-BSA. Salbutamol immunogen solution is referred to as SAL-BSA solution.
[0057] After diluting the SAL-BSA solution with PBS buffer, measure the spectrophotometric values at 280nm and 260nm, calculate the protein concentration in the dilution according to the formula, and mul...
Embodiment 3
[0071] Embodiment 3, the preparation of albuterol monoclonal antibody
[0072] Balb / c mice: purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.;
[0073] SP2 / 0 myeloma cells: purchased from Sigma-Aldrich Company, the product catalog number is 08060101.
[0074] 1. Animal immunity
[0075] The SAL-BSA solution prepared in Example 2 was used to immunize Balb / c mice. Each mouse was immunized with 100 μg of SAL-BSA once, and immunized 4 times in total, with an interval of two weeks between each time. Injection, the last immunization way is intraperitoneal injection.
[0076] 2. Cell fusion and cloning
[0077] 1. Three days after the fourth immunization, splenocytes were collected and fused with SP2 / 0 myeloma cells at a ratio of 5:1 (quantity ratio). Cell supernatants were measured by indirect competitive ELISA, and positive wells were screened.
[0078] 2. The positive wells were cloned by the limiting dilution method to obtain a hybridoma cell capab...
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