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Salbutamol immunomagnetic bead separation and enrichment kit and application thereof

An albuterol, separation and enrichment technology, applied in the field of immunology, can solve the problems of high binding strength, low binding probability of antibodies and detection target substances, etc., and achieves time saving, environmental protection, high specificity and accuracy of washing and elution processes. sexual effect

Active Publication Date: 2017-07-07
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The covalent bond between the antibody and the carrier is a physical binding force such as hydrophobic force and ion exchange force, and its binding force is not as strong as that of the chemical bond of immunomagnetic beads
Moreover, the carrier of the immunoaffinity column is a solid phase, and the binding probability of the antibody to the detection target substance will be lower than that of the immunomagnetic beads.

Method used

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  • Salbutamol immunomagnetic bead separation and enrichment kit and application thereof
  • Salbutamol immunomagnetic bead separation and enrichment kit and application thereof
  • Salbutamol immunomagnetic bead separation and enrichment kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Preparation of specific components of the kit

[0021] 1. Albuterol Hapten Synthesis

[0022] The albuterol hapten is produced by diazotization reaction of albuterol and p-aminobenzoic acid, and a spacer arm with a carboxyl group is introduced next to the phenolic hydroxyl group to obtain the albuterol hapten.

[0023] Take 0.5g of p-aminobenzoic acid, add it to 300μL of 1mol / L hydrochloric acid solution, add 4mL of distilled water to dilute, cool to 0°C and stir, add 0.23g, continue stirring for 2h to obtain a diazonium salt solution. Take 0.7g salbutamol, dissolve it with methanol, add 0.21g sodium hydroxide to it, stir, add diazonium salt solution, continue stirring for 4h, stop the reaction, add dilute hydrochloric acid to adjust the pH value to 5.0, add ethyl acetate to extract, separate For the aqueous phase, the organic phase was evaporated to dryness, and diethyl ether was added for recrystallization to obtain 0.61 g of the albuterol hapten produc...

Embodiment 2

[0036] Embodiment 2: the formation of kit

[0037] Set up detection albuterol magnetic immunomagnetic bead separation and enrichment kit so that it contains the following components:

[0038] Magnetic Beads Conjugated to Salbutamol Monoclonal Antibody

[0039] Reconstitution solution

[0040] magnet

[0041] Add the magnetic beads coupled with albuterol monoclonal antibody to the reconstitution solution to a final concentration of 10 mg / mL.

Embodiment 3

[0042] Example 3: Kit for separation and enrichment of salbutamol in samples

[0043] 1) Take 0.1 mL of magnetic bead reconstitution solution conjugated with albuterol monoclonal antibody in a 10 mL centrifuge tube, wash the magnetic beads with 5 mL of deionized water for 1-2 times, and place the centrifuge tube on the magnet for 2-3 minutes. Make sure that the magnetic beads are all adsorbed on the magnet, and separate the magnetic beads and washing solution with a magnet every time;

[0044] 2) Add 5 mL of urine sample into a 10 mL centrifuge tube filled with rinsed magnetic beads coupled with salbutamol monoclonal antibody, mix well, and react at room temperature for 20 min; or homogenize animal tissue and feed samples with a homogenizer After quality, weigh 5.0±0.05g sample into the sample bottle, add 1.5±0.05g sodium chloride and 20mL50% methanol solution respectively, vortex with vortex for 5min, centrifuge at 3000r / min for 5min at room temperature, or after standing Ta...

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Abstract

The invention relates to a salbutamol immunomagnetic bead separation and enrichment kit which comprises magnetic beads, reconstitution fluid and a magnet, wherein the magnetic beads are coupled with a salbutamol monoclonal antibody; the magnetic beads coupled with the salbutamol monoclonal antibody are prepared by mixing the salbutamol monoclonal antibody with magnetic beads in a mass ratio of 1:(500-600) and dissolving in 2-morpholinoethanesulfonic acid, monohydrate through coupling; the salbutamol monoclonal antibody is prepared by immunizing Balb / c mice by using conjugate of salbutamol hapten and bovine serum albumin as immunogen; the salbutamol hapten is prepared by performing diazo-reaction on salbutamol and aminobenzoic acid and by introducing carboxyl-containing spacers beside phenolic hydroxyl groups. The invention further discloses a sample pretreatment method for separating salbutamol by using the salbutamol immunomagnetic bead separation and enrichment kit. The method has relatively high specificity for salbutamol and is high in recycling rate and accuracy, sample pretreatment steps are simplified, and multiple and massive organic solvents are not used in the sample pretreatment process.

Description

technical field [0001] The invention relates to a salbutamol immunomagnetic bead kit and a method for separating and enriching salbutamol in a sample. in the field of immunology. Background technique [0002] Salbutamol (Salbutamol, SAL), also known as albuterol, is a selective β-receptor agonist, which can selectively excite the β-receptors of bronchial smooth muscle, and has a strong bronchodilator effect. For the prevention of bronchial asthma. Experiments have confirmed that SAL can promote animal skeletal muscle growth and reduce fat accumulation. Humans will experience poisoning symptoms such as muscle tremors, myalgia, headache, nausea, and vomiting after eating animal tissues with high salbutamol content. After the illegal use of clenbuterol hydrochloride was greatly restricted, lawbreakers had to find new substitutes, such as albuterol and cimaterol. Therefore, some farms at home and abroad illegally use this class of drugs. The "List of Veterinary Drugs and Ot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 罗晓琴朱亮亮刘玉梅冯月君赵正苗王琳琛徐念琴
Owner BEIJING KWINBON BIOTECH
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