Preparation and application of miniature high-efficiency clenbuterol immuno-affinity chromatography column

Abstract

The invention relates to a preparation method and application of a miniature high-efficiency clenbuterol immuno-affinity chromatography column. The technical scheme is characterized by comprising the following steps of: preparing a high-quality antibody, synthesizing efficient prepared bromoacetyl chloride clenbuterol purified by a liquid chromatogram and thiolation hemocyanin into immunogen immune animals, and obtaining antiserum; reacting the purified bromoacetyl chloride clenbuterol with thiolation agarose to obtain clenbuterol-agarose padding, loading the padding into an antigen affinity chromatography column; and purifying a clenbuterol specific antibody in the antiserum by using the antigen affinity chromatography column. The preparation method of the clenbuterol immuno affinity chromatography column comprises the following steps of: coupling high-quality antibodies at high density to the agarose oxidized by periodic acid to prepare an antibody affinity padding, and placing 25 mul of padding in a small specially-made column to prepare the miniature immuno-affinity chromatography column. The immuno-affinity chromatography column provided by the invention can be used for specifically gathering clenbuterol in a sample to be tested at high efficiency and can increase the accuracy, reliability and sensitiveness of detection when being combined with an analytic instrument and colloidal gold test paper for use.

Description

technical field [0001] The invention relates to a biological separation and enrichment technology, in particular to a preparation technology of a miniature high-efficiency clenbuterol immunoaffinity chromatography column and its application. Background technique [0002] Clenbuterol hydrochloride is a β-stimulant, which can increase lean meat percentage, reduce fat deposition and promote animal growth. It is widely used as a growth promoter by farmers or breeding enterprises, resulting in a large amount of clenbuterol hydrochloride in animal products. residual. Human consumption of such animal products can cause muscle tremors, heart palpitations, nervousness, headache, dizziness, nausea, vomiting, fever, tremors and other symptoms, seriously endangering the health of consumers. Therefore, many countries (including my country) have now been banned from adding clenbuterol to feed to increase the lean meat percentage of animals, and the content of clenbuterol in livestock pro...

AI Technical Summary

Problems solved by technology

[0010] 4) There is no miniaturized affinity column

The above-mentioned reasons lead to poor enrichment efficiency and purification ability of clenbuterol in the sample to be tested by the prepared affinity chromatographic column, resulting in shortcomings such as poor detection sensitivity, low accuracy, and low reliability.

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