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Method for purifying halofuginone and its special immune affinity chromatographic column

A halofuginone and immunoaffinity technology, applied in the field of immunoaffinity chromatographic column, can solve the problems of excessive organic solvent waste, no IAC column for sale, cumbersome processing process, etc., to improve the analysis quality, save the organic solvent, and have a good purification effect. Effect

Inactive Publication Date: 2006-07-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pretreatment of these methods utilizes liquid-liquid distribution and conventional SPE column purification and separation, all of which have disadvantages such as cumbersome treatment process, poor purification effect, waste of organic solvents, and long time required to varying degrees.
Immunoaffinity technology is a new technology applied in the field of analysis in the 1990s, but there is no report on the purification of hemosanone in the matrix with an immunoaffinity column, and there is no commercial IAC column for sale

Method used

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  • Method for purifying halofuginone and its special immune affinity chromatographic column

Examples

Experimental program
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Embodiment 1

[0025] Embodiment 1, the preparation of the immunochromatographic column of purifying hemosanone

[0026] 1. Preparation of hemosanone rabbit polyclonal antibody

[0027] Synthesis of hiradhenone hapten: dissolve hiradhenone hydrobromide (HAL) in N,-N, in dimethylformamide (DMF), add N-trimethylsilyl imidazole (TMSI, N-Trimethysiylimidazole), The resulting white precipitate is a derivative of hemosanone succinic acid, which is a hemosanone hapten.

[0028] Preparation of the immunogen: Using N-hydroxysuccinic acid imide (activated lipid method), the hemosanone hapten is coupled with the protein carrier bovine serum albumin (BSA) or ovalbumin (OVA) to prepare the immunogen BSA- HAL or OVA-HAL.

[0029] Animal immunization: New Zealand white rabbits are used as immunized animals, and the immunization dose is 1mg·ml -1 BSA-HAL conjugate or 1mg·ml -1 OVA-HAL conjugates. New Zealand rabbits were bred for several weeks and then immunized. The first immunization was emulsified w...

Embodiment 2

[0049] Example 2. Preparation of kits for immunoaffinity chromatographic columns coupled with rabbit polyclonal antibody IgG or mouse monoclonal antibody and its purification effect on hemosanone

[0050] 1. Preparation of a kit for purifying hemosanone

[0051] It is mainly composed of a box body, an immunoaffinity chromatographic column (IAC column), HAL standard solution, diluent, washing solution, eluent, preservation solution, and a sponge bracket. The sponge bracket is provided with holes and grooves. In the groove of the sponge bracket, there are reagent bottles containing HAL standard solution, diluent, washing solution, eluent, and preservation solution, and the hole of the sponge bracket is equipped with an IAC column. The immunochromatographic column is the immunoaffinity column prepared in Example 1 coupled with the monoclonal antibody secreted by the monoclonal hybridoma cell line A-2--2CGMCC No.1608 coupled with rabbit polyclonal antibody IgG or hemosanone.

[0...

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Abstract

The invention discloses a method of dichroa febrifuga ketone's purification and its immune affinity chromatographic column. The immune affinity adsorption chromatography column of puriftying dichroa febrifuga ketone is loaded with adsorbent which is consisted of solid carrier and dichroa febrifuga ketone polyclonal antibody dichroa or monoclonal antibody connected with the solid carrier; the dichroa febrifuga ketone polyclonal antibody dichroa or monoclonal antibody is obtained by the immunogen of connective thing of dichroa febrifuga ketone haptene and carrier protein; the dichroa febrifuga ketone haptene is made by dissolving hydrobromic acid dichroa febrifuga ketone in N, -N, dimethyl formamide, then adding trimethylchlsilane imidazole, the outcome dichroa febrifuga ketone succinic derivant is dichroa febrifuga ketone haptene. The purifying method of this invention combining chromatography can examine the content of dichroa febrifuga ketone efficiently. This method covers the shortage of measuring sample directly only by immunoassay technology which gets little information, bad quantitative accuracy or insufficient selective of physicochemical method, incarnates the complementarity of immunology technic and general physicochemical technic in the analytic mechanism.

Description

technical field [0001] The invention relates to a method for purifying hemosanone and a special immunoaffinity chromatographic column thereof. Background technique [0002] With the development of life sciences, people have become more and more interested in the substances and their changes in living organisms, and the analysis of biological samples has become a necessary means to explore and discover the mysteries of life. Due to the complex components of biological samples, the concentration of analytes is low, and most of the sample volumes are small, which puts forward higher requirements for the selectivity and sensitivity of the analytical method. Immunoaffinity chromatography (IAC, immunoaffinity chromatography) is an analytical method that combines an immune response with a chromatographic analysis method. Its high selectivity and high affinity undoubtedly simplify the analysis process. In the analysis of veterinary drug residues, the simplest and most effective ap...

Claims

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Application Information

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IPC IPC(8): C07D401/06G01N33/53G01N33/559
Inventor 沈建忠丁双阳史为民侯亚莉何方洋
Owner CHINA AGRI UNIV
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