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
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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.
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Abstract
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