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Kitasamycin magnetic immunochemiluminescence detection kit

A chemiluminescence detection and kitasamycin technology, which is applied in measuring devices, scientific instruments, instruments, etc., can solve the problems of complex processing, time-consuming, unsuitable for instant detection, etc., and achieve the effect of high sensitivity

Inactive Publication Date: 2018-02-09
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the advantages of accuracy, stability, and good specificity, and can be used as standard methods, but the instruments and equipment are expensive, heavy, require a large amount of solvents, and have high requirements for operators. The sample pretreatment is complicated, time-consuming, laborious, and not easy to popularize. Point-of-care testing of large-scale samples
Although microbial detection methods can perform real-time detection of a large number of samples, they have poor specificity and cannot perform accurate qualitative and quantitative analysis.

Method used

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  • Kitasamycin magnetic immunochemiluminescence detection kit
  • Kitasamycin magnetic immunochemiluminescence detection kit
  • Kitasamycin magnetic immunochemiluminescence detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The preparation of embodiment 1 kitasarmycin hapten and immunogen

[0027] Weigh 100 mg of kitasarmycin standard substance and dissolve in 50 mL of methanol. Weigh 50 mg of oxycarboxymethyl amine and dissolve in 10 mL of water. The oxygen carboxymethyl amine solution was slowly added to the kitasamycin methanol solution, and the reaction was stirred overnight. The reaction solution was rotary evaporated to dryness at 60°C. Accurately draw 10 mL of DMF to dissolve the evaporated matter, add 1 g of dicyclohexylcarbodiimide and stir overnight to obtain the kitasarmycin hapten.

[0028] Weigh 35 mg of carbonyldiimidazole, fully dissolve it with 0.7 ml of acetone, add 30 mg of kitasarmycin hapten and shake it at 37°C for 2 hours, then dry it under vacuum overnight; Shake at 4°C for 3.5 days; dialyze with 0.2mol / L boric acid buffer solution of pH 8.0 for 2 days, add thimerosal, and store at 4°C; use ultraviolet scanning method to identify and confirm that the conjugate is ...

Embodiment 2

[0029] Preparation of embodiment 2 enzyme-labeled antigen

[0030] Take 25mg kitasarmycin hapten, dissolve in 1.2mL N,N-dimethylformamide (DMF); take 15mg dichloroethane (EDC) and 15mg N-hydroxysuccinimide (NHS) with 0.4mL After the water is fully dissolved, add it to the hapten solution and stir at room temperature for 24 hours to obtain the reaction solution A; weigh 50 mg of horseradish peroxidase (HRP) and fully dissolve it in pH 7.2, 5 mL of phosphate buffered saline In the solution, the reaction solution A was slowly added dropwise to the HRP solution, and stirred at room temperature for 24 hours; dialyzed with 0.01mol / L phosphate buffer solution at 4°C for 4 days, and the dialysate was changed 3 times a day to remove Unreacted small molecule substances were obtained from kitasamycin enzyme-labeled antigen; aliquoted and stored at -20°C for future use.

Embodiment 3

[0031] Embodiment 3 Preparation of kitasarmycin monoclonal antibody

[0032] Animal immunization: use the immunogen prepared in Example 1 at 120 μg / mouse, dissolve the immunogen in physiological saline and mix it with Freund's complete adjuvant in an equal volume, and inject subcutaneously on the back of the neck to immunize 6-8 week-old Balb / c female mice. On the 7th, 14th, and 28th day after the initial immunization, mix the immunogen and Freund's incomplete adjuvant in equal volumes, and add immunization once each, and add 120 μg of the immune complex per mouse 3 days before the fusion, without Freund's adjuvant. Immune once.

[0033] Cell fusion: according to the conventional method, take the splenocytes of the immunized mice and mix them with the mouse myeloma cells (SP2 / 0) in the logarithmic growth phase, and then slowly add the preheated fusion agent (PEG4000) within 45 seconds. Fusion, suspend evenly with HAT medium, then add appropriate amount of feeder cells, cultur...

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Abstract

The invention relates to a high sensitivity, high accuracy and high accuracy kitasamycin magnetic immunochemiluminescence detection kit. The kit comprises a horse radish peroxidase-labeled kitasamycinhapten label, an enzyme-labeled antigen diluent, a magnetic label antibody, a magnetic label antibody diluent, a series of kitasamycin standard solutions, a concentrated compound solution, a concentrated cleaning solution and solutions A and B of chemiluminescent substrates. A lot of tests screen specific monoclonal antibodies with excellent properties and determine appropriate concentrations ofcomponents in the reaction solution. The kitasamycin magnetic immunochemiluminescence detection kit can fast and accurately detect low content kitasamycin and is suitable for detection of kitasamycinresidue content of pig or poultry animal foods.

Description

technical field [0001] The invention relates to a chemiluminescence detection kit and a detection method thereof, in particular to a magnetic immunochemiluminescence detection kit and a method for measuring the residual amount of kitasamycin in a sample to be tested by using the kit. Background technique [0002] With the improvement of people's quality of life, food safety issues are more and more concerned by consumers, especially the problem of drug residues such as pesticides and antibiotics is the focus of consumers' attention. On food safety, a hot issue of global concern, how to quickly and accurately detect food safety has become a top priority. To detect antibiotic residues in food, it is necessary to choose a detection method with strong specificity, high sensitivity, high efficiency and rapidity. Therefore, it is of great significance to study methods for the detection of various antibiotic residues and improve their sensitivity and selectivity. [0003] Antibio...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/532
CPCG01N33/535G01N33/532G01N2333/36
Inventor 谷正赵凯刘芳许晨李健李云龙
Owner QINGDAO UNIV
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