Immunoaffinity chromatographic column adopting astragaloside IV as ligand and application of immunoaffinity chromatographic column

Abstract

The invention discloses a technical scheme of an immunoaffinity chromatographic column adopting astragaloside IV as a ligand and an application of the immunoaffinity chromatographic column. The immunoaffinity chromatographic column adopts the astragaloside IV as the ligand and is prepared through specific steps as follows: an astragaloside IV hapten is prepared; an immunoaffinity adsorbent is prepared from the astragaloside IV hapten and a solid-phase carrier through cross-linking; the immunoaffinity adsorbent is taken as packing to be charged into a chromatographic column; the packing is washed alternately with an acetic acid buffer solution and a Tris-HCl buffer solution for three times, and the immunoaffinity chromatographic column is obtained. A method adopting the prepared immunoaffinity chromatographic column to purify an astragaloside IV antibody comprises steps as follows: a to-be-purified sample is added to the immunoaffinity chromatographic column and is eluted by an eluent and neutralized by a neutralization solution and the like. According to the invention, the purification efficiency of the astragaloside IV monoclonal antibody is effectively improved, the purity can be higher than 99%, besides, the operation is simple, the cost is low, and the pollution is low.

Description

technical field [0001] The invention relates to an immunoaffinity chromatography column using astragaloside IV as a ligand and application thereof. Background technique [0002] Astragalus is a commonly used traditional Chinese medicine, which has the functions of invigorating qi and strengthening the surface, suppressing sores and promoting granulation, diuresis and detoxification. Modern pharmacological studies have shown that Astragalus has various pharmacological activities such as enhancing the body's immunity, lowering blood sugar, strengthening the heart, diuresis, lowering blood pressure, anti-aging and anti-fatigue. The main active ingredients in Astragalus are astragalus polysaccharides, astragalus saponins and isoflavones. At present, Astragaloside IV in Astragalus saponins is mainly used as the standard for evaluating the quality of Astragalus medicinal materials. The current methods for quantitative analysis of astragaloside mainly include: high performance liq...

AI Technical Summary

Problems solved by technology

At present, the antibody purification methods are mainly salting-out method, organic solvent precipitation method, chromatography of various principles, and affinity purification method (usually ProteinA or ProteinG), etc., all of which cannot obtain very pure specific antibodies. And purification method, can only get the mixture of specific antibody and non-specific antibody, but can not separate the two to get specific antibody, which affects the accuracy and efficiency of evaluating the quality of Radix Astragali. A purification method for effectively separating and purifying specific antibodies, with high academic value and application prospects

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