Immunoaffinity column for quickly purifying and enriching cimaterol as well as preparation method and application of immunoaffinity column

A cimaterol, immunophilic technology, applied in the field of immunological separation, purification and enrichment, to improve the accuracy and reliability of detection, high antibody specificity, and eliminate the interference of impurities

Inactive Publication Date: 2014-04-09
NANNING LANGUANG BLUE LIGHT BIOTECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the detection of cimaterol by immunoaffinity chromatography. Therefore, there is an urgent need to develop an immunoaffinity chromatography detection technology for cimaterol.

Method used

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Examples

Experimental program
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Embodiment

[0022] Example Preparation of Immunoaffinity Column for Rapid Purification and Enrichment of Cimaterol

[0023] 1. The cimaterol is diazotized by the diazo method and then coupled to the carrier protein to obtain an immunogen, and the immunogen is immunized with animals to produce antiserum; the specific steps are as follows:

[0024] (1) Preparation of cimaterol immunogen

[0025] ① Weigh 10mg of cimaterol standard, dissolve in 100uL tetrahydrofuran, add 900μL 1mol / L HCl, freeze for 5 minutes; add 50mg solid sodium nitrite, shake well after freezing for 2 minutes, continue to react at 4°C for 30 minutes, and obtain the weight Nitrided cimatero;

[0026] ②Weigh 100mg of bovine serum albumin and dissolve it in 5mL0.1mol / LNa 2 CO 3 , freeze for 2 minutes and then add 0.5 mL of saturated Na 2 CO 3 ;

[0027] ③ Add the cimaterol obtained in step ① to the bovine serum albumin solution in step ② and react overnight on a shaker at room temperature; desalt with SephadexG25 to ob...

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PUM

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Abstract

The invention discloses an immunoaffinity column for quickly purifying and enriching cimaterol. The immunoaffinity column is obtained by loading cimaterol-resistant immunoaffinity filler into an empty column tube; the cimaterol-resistant immunoaffinity filler consists of a solid phase carrier and a cimaterol specificity polyclonal antibody coupled with the solid phase carrier; the cimaterol specificity polyclonal antibody is obtained by preparing antiserum through a conjugate which is obtained by coupling the cimaterol diazotized by a diazotizing method with a carrier protein and then purifying the antiserum by an immunoaffinity method. The invention also discloses a preparation method and application of the immunoaffinity column. The specificity of the antibody used by the immunoaffinity column disclosed by the invention is high; the immunoaffinity column disclosed by the invention is used for pretreating a sample; then the immunoaffinity column is used after being combined with a chromatographic method and a colloidal gold test strip, so that the content of the cimaterol can be efficiently and accurately detected.

Description

technical field [0001] The invention belongs to the technical field of immunological separation, purification and enrichment, and relates to an immunoaffinity column for rapid purification and enrichment of cimaterol, a preparation method and application thereof. Background technique [0002] Cimaterol (CIM) is a kind of phenylethylamine drugs, which is a β-stimulant. It is mainly used in medicine and veterinary medicine to dilate airways and increase lung ventilation. It can be used for asthma, obstructive pneumonia, smooth muscle convulsions and shock embolism. Studies have shown that CIM can increase lean meat percentage and promote animal growth. Because of its obvious economic benefits, farmers or breeding enterprises are motivated to use it in large quantities, resulting in a large amount of cimaterol residues in animal products. Human consumption of such animal products can cause muscle tremors, heart palpitations, nervousness, headache, dizziness, nausea, etc. Pois...

Claims

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Application Information

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IPC IPC(8): B01J20/22B01J20/30B01D15/38C07K16/44C07K16/06C07K17/10G01N1/34G01N1/40
Inventor 谢体三潘丽金宁欢欢刘晓松吕春秋张兰叶开富萧洁
Owner NANNING LANGUANG BLUE LIGHT BIOTECH
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