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Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent

A technology for docetaxel and detection reagents, applied in biological testing, immunoglobulin, immunoglobulin from serum, etc., can solve the problems that cannot meet the needs of clinical testing, high-throughput and low-cost clinical needs, etc. Achieve effects that are conducive to individualized treatment, reduce testing costs, and have strong specificity

Active Publication Date: 2019-02-26
苏州博源医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the main methods for monitoring the plasma concentration of docetaxel at home and abroad are enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), reversed-phase high-performance liquid chromatography (RP-HPLC), liquid chromatography- Traditional methods such as tandem mass spectrometry (LC-MS / MS), but these methods cannot meet the clinical needs of high throughput and low cost
Although the docetaxel assay kit that uses latex-enhanced immune turbidimetry and can be applied to biochemical analyzers has been launched in China, it is far from meeting the growing demand for clinical testing.
At present, there is a lack of docetaxel detection reagents with good stability, high sensitivity, and strong specificity in the market, especially high-quality automated detection reagents. Docetaxel determination reagents for automatic biochemical analyzers have become a hot spot in the in vitro diagnostic reagent industry at home and abroad

Method used

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  • Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent
  • Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent
  • Docetaxel immunogen, anti-docetaxel specific antibody and docetaxel detection reagent

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a docetaxel immunogen, a synthetic method of the docetaxel immunogen, an anti-docetaxel antibody obtained by the immunogen, a detection reagent and a preparation method of the anti-docetaxel antibody. The docetaxel immunogen disclosed by the invention is synthesized by brand-new docetaxel derivatives and has the advantage of high immunogenicity, an anti-docetaxel specific antibody with high specificity and binding force can be obtained by induction, and any cross reaction with common 62 medicines is avoided. According to the docetaxel detection reagent prepared from the antibody, the content of docetaxel in a sample can be conveniently and accurately determined, and clinical individualized treatment is promoted. Compared with the method in the prior art, the detection reagent disclosed by the invention can realize high-flux and rapid detection on a fully automatic biochemical analyzer and has the advantages of simple and convenient operation, high sensitivity, high specificity, accurate result and the like, the detection cost of the docetaxel can be effectively reduced, and the detection reagent is suitable for large-scale clinical popularization and application.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a docetaxel immunogen, an anti-docetaxel specific antibody and a docetaxel detection reagent. Background technique [0002] Docetaxel (Docetaxel) structural formula is shown in formula (Ⅲ): [0003] [0004] Docetaxel is a new type of semi-synthetic antineoplastic drug, which belongs to paclitaxel compounds. Its mechanism of action is to strengthen tubulin polymerization and inhibit microtubule depolymerization, resulting in the formation of stable non-functional microtubule bundles, thereby destroying tumor cells of mitosis. In recent years, docetaxel has been widely used in the clinical treatment of breast cancer, advanced non-small cell lung cancer, gastric cancer and other cancers, but its adverse reactions are still relatively obvious, mainly including: bone marrow suppression, allergic reactions, skin reactions, and other adverse reactions Reactions also include nausea, vomi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/765C07K14/795C07K14/47C07K1/107C07K16/44C07K16/06G01N33/543G01N33/531
CPCC07K1/1077C07K14/47C07K14/765C07K14/795C07K16/06C07K16/44C07K19/00G01N33/531G01N33/68G01N33/94
Inventor 虞留明涂建成卢忠心王英徐琦鋆
Owner 苏州博源医疗科技有限公司
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