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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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 re

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

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[0025] Example 1. Preparation of immunochromatographic column for purifying changshanone

[0026] 1. Preparation of rabbit polyclonal antibody of Changshanone

[0027] Synthesis of changshanone hapten: Dissolve changshanone hydrobromide (HAL) in N, -N, dimethylformamide (DMF), add N-trimethylsilyl imidazole (TMSI, N-Trimethysiylimidazole), The white precipitate produced is the changshanone succinic acid derivative, which is the changshanone hapten.

[0028] Preparation of immunogen: Using N-hydroxysuccinic acid imine (active lipid method), the changshanone hapten is coupled with the protein carrier bovine serum albumin (BSA) or ovalbumin (OVA) to make 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 conjugate. New Zealand rabbits were raised for several weeks and then vaccinated. The first immunization was emulsified with 1ml comple...

Example Embodiment

[0049] Example 2. Preparation of a kit of immunoaffinity chromatography column coupled with rabbit polyclonal antibody IgG or mouse monoclonal antibody and its purification effect on changshanone

[0050] 1. Preparation of kit for purifying changshanone

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

[0052] Among them, ...

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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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