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Method for preparing clenbuterol liquid phase chip probe

A liquid phase chip and probe technology, applied in the field of immunochemistry, can solve the problems of poor reaction repeatability, limited application and promotion, etc., and achieve the effects of good stability, strong controllability and high yield

Inactive Publication Date: 2011-02-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This process uses the carrier protein as the linker arm, and its practical application is limited due to the poor reproducibility of the coupling reaction between the carrier protein and the small molecule and the reaction between the conjugate and the microsphere.

Method used

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  • Method for preparing clenbuterol liquid phase chip probe
  • Method for preparing clenbuterol liquid phase chip probe
  • Method for preparing clenbuterol liquid phase chip probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 The principle of the preparation method of the clenbuterol liquid phase chip probe is shown in the following formula:

[0023]

[0024] In the above formula, ● is carboxylated microspheres, Sulfo-NHS is nitrogen hydroxysuccinimide sulfonate, and EDC is water-soluble carbodiimide.

[0025] The method comprises the following specific steps:

[0026] (1) Surface-modified p-aminobenzoic acid linker of fluorescent microspheres

[0027] Take 5×10 5 Carboxylated polystyrene fluorescent microspheres, washed with water, centrifuged and removed the supernatant, added 100 μL activation solution (0.1M phosphate buffer solution, pH 6.0), vortexed for 1min, then ultrasonicated for 1min, added freshly prepared 50mg / ml nitrogen hydroxysuccinimide sulfonate (sulfo-NHS) and water-soluble carbodiimide (EDC. Centrifuge the ball to remove the supernatant, add 0.5mL of 0.01wt% p-aminobenzoic acid solution, incubate in the dark at room temperature for 60min, remove the supern...

Embodiment 2

[0033] Example 2 The principle of the preparation method of the clenbuterol liquid phase chip probe is shown in the following formula:

[0034]

[0035] In the above formula, ● is carboxylated microspheres, Sulfo-NHS is nitrogen hydroxysuccinimide sulfonate, and EDC is water-soluble carbodiimide.

[0036] The method is as follows:

[0037] (1) Surface-modified m-aminobenzoic acid linking arm of fluorescent microspheres

[0038] take 10 6 Carboxylated polystyrene fluorescent microspheres were washed with water, centrifuged and the supernatant was removed, then 100 μL of activation solution (0.05M phosphate buffer solution, pH 6.5) was added, vortexed for 1 min, and then ultrasonicated for 3 min. Add 10 μL each of freshly prepared 50 mg / ml nitrogen hydroxysuccinimide sulfonate (sulfo-NHS) and water-soluble carbodiimide (EDC.HCl) aqueous solutions, vortex and mix for 10 seconds, and incubate with shaking at room temperature for 20 minutes in the dark. Centrifuge the activat...

Embodiment 3

[0043] Embodiment 3 The principle of the preparation method of the clenbuterol liquid phase chip probe is shown in the following formula:

[0044]

[0045] In the above formula, ● is carboxylated microspheres, Sulfo-NHS is nitrogen hydroxysuccinimide sulfonate, and EDC is water-soluble carbodiimide.

[0046] The method comprises the steps of:

[0047] (1) Surface-modified o-aminobenzoic acid linker of fluorescent microspheres

[0048] Take 2.5×10 6 Carboxylated polystyrene fluorescent microspheres were washed with water, centrifuged and the supernatant was removed, then 100 μL of activation solution (0.01M phosphate buffer solution, pH 7.0) was added, vortexed for 1 min, and then sonicated for 5 min. Add 10 μL each of freshly prepared 50 mg / ml nitrogen hydroxysuccinimide sulfonate (sulfo-NHS) and water-soluble carbodiimide (EDC.HCl) aqueous solutions, vortex and mix for 10 seconds, and incubate for 60 minutes at room temperature in the dark with shaking. The activated mi...

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Abstract

The invention relates to a method for preparing a clenbuterol liquid phase chip probe in the technical field of immunochemistry. The method comprises the following steps of: modifying the surface of a carboxylated polystyrene fluorescent microsphere with p-aminobenzoic acid, m-aminobenzoic acid or o-aminobenzoic acid by adopting a carbodiimide method; and then marking clenbuterol molecules on the surface of the fluorescent microsphere by the carbodiimide method to finally obtain the clenbuterol liquid phase chip probe. The method for preparing the clenbuterol liquid phase chip probe has the advantages of simple operation, high stability and favorable clenbuterol targeted property of products and can provide a more convenient approach for the development of test technology of the clenbuterol liquid phase chip.

Description

technical field [0001] The invention relates to a preparation method of a clenbuterol detection reagent in the technical field of immunochemistry, in particular to a preparation method of a clenbuterol liquid phase chip probe. Background technique [0002] Clenbuterol (Clenbuterol, CL) is a synthetic β2-receptor hormone, which can increase the lean meat percentage of several livestock including pigs after being added to the feed. It is well absorbed in the body. After eating by pigs, it promotes protein synthesis in the metabolic process, accelerates the conversion and decomposition of fat, and improves the lean meat rate of pork, so it is called lean meat powder. Its bioavailability is so high compared to other beta-agonists that consumption of pork containing clenbuterol has been toxic. Human consumption of pig liver or pig lung is enough to cause poisoning. There is no formal organization in the world that approves clenbuterol as a feed additive for animal growth promot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F8/30C08F12/08C08F8/32G01N33/533
Inventor 彭池方匡华陈伟刘丽强胥传来
Owner JIANGNAN UNIV