Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent
A docetaxel and immunogen technology, applied in the biological field, can solve the problems of not being able to meet the clinical needs of high-throughput and low cost, as well as the needs of clinical testing, so as to facilitate individualized treatment, reduce testing costs, and improve results. accurate effect
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Embodiment 1
[0067] Example 1: Synthesis and quantitative detection of docetaxel derivatives
[0068] The chemical structure of docetaxel derivatives is shown in formula (IV):
[0069]
[0070] The synthetic route and preparation steps of the above-mentioned docetaxel derivatives are as follows:
[0071]
[0072] 1. Weigh 1g of docetaxel and 2g of succinic anhydride into the reaction system, and then incubate with 20ml of pyridine or dry benzene in a glass flask continuously refluxing.
[0073] 2. The reaction mixture was continuously heated at a reflux temperature of 70-80° C. for 6 hours, then the reaction mixture was slowly cooled to room temperature, and excess pyridine or benzene was poured off gently.
[0074] 3. The remaining organic components are continuously fed with a nitrogen flow under negative pressure to be evaporated and dried, and the obtained dry residual product is the docetaxel derivative.
[0075] 4. The product obtained above was washed with 60% ethanol prepar...
Embodiment 2
[0077] Example 2: Synthesis of Docetaxel Immunogen
[0078] Docetaxel immunogen is composed of bovine serum albumin (Bovine Serum Albumin, BSA) and -CO-(CH 2 ) n The -COO- group is connected. In this embodiment, the synthesis method of the immunogen is described in detail by taking n=2 as an example. The specific steps are as follows:
[0079] 1. Dissolve 200mg of BSA in 50ml of 0.2M, pH 8.5 phosphate buffer;
[0080] 2. Add the following chemicals into a small beaker and stir to dissolve at room temperature for 30 minutes: 200mg of docetaxel derivatives, 3.5ml of dimethylformamide, 3.5ml of ethanol, 7.0ml of 10mM potassium phosphate buffer at pH 5.0, 200mg 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide, 50mg N-hydroxysulfosuccinimide;
[0081] 3. The dissolved solution was added dropwise to the BSA solution, and stirred overnight at 2-8° C. to obtain the antigen; the synthesized antigen was purified by dialysis to obtain the docetaxel immunogen.
Embodiment 3
[0082] Example 3: Preparation of Anti-Docetaxel Specific Antibody
[0083] The docetaxel immunogen prepared in Example 2 was inoculated into experimental animal rabbits by a conventional method, and the antiserum was taken after booster immunization, and the specific steps were as follows:
[0084] 1. Dilute the synthetic docetaxel immunogen to 1.0 mg / ml with PBS to obtain an antigen solution, then mix 1.0 ml of the antigen solution with Freund's complete adjuvant, and inject the experimental animal rabbit.
[0085] 2. After 2-3 weeks, inject 1.0ml of the same antigen solution and incomplete Freund's adjuvant to the above-mentioned experimental animal rabbit once, and then inject once every four weeks, a total of 4 injections.
[0086] 3. Take blood from the experimental animal rabbit in step 2, separate and purify to obtain anti-docetaxel specific antibody with a titer of 1:30000-1:50000.
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