Diarrhetic shellfish poison immunoaffinity column, preparation method and application thereof

Abstract

The invention belongs to the technical field of food safety inspection, and specifically relates to a diarrhetic shellfish poison immunoaffinity column, a preparation method and application thereof. The preparation method comprises the following steps: firstly using agarose gel activated by using an epoxy chloropropane activation method as a carrier, then coupling gene recombinant protein G onto the carrier to obtain protein G-sepharose, then connecting a diarrhetic shellfish poison antibody onto the protein G-sepharose so as to obtain antibody-protein G-sepharose, then using a cross-linking agent for cross-linking so as to obtain an antibody-protein G-sepharose filler, and finally packing the antibody-protein G-sepharose filler so as to form the diarrhetic shellfish poison immunoaffinitycolumn with high purity and affinity. The specific binding characteristic of the gene recombinant protein G and the diarrhetic shellfish poison antibody is sufficiently utilized, and the Fab segment of the diarrhetic shellfish poison antibody is exposed outside, so that the antibody combining capacity of the gene recombinant protein G, the capturing capacity of the diarrhetic shellfish poison andthe purification efficiency of the diarrhetic shellfish poison are greatly improved.

Description

Technical field: [0001] The invention belongs to the technical field of food safety detection, and in particular relates to a diarrhea shellfish toxin immunoaffinity column and a preparation method and application thereof. Background technique: [0002] The affinity column is a column installed after the substance that has a specific binding effect with the purification object is connected to an insoluble carrier to make an affinity adsorbent, and it is mostly used for affinity chromatography. [0003] Some red tide organisms that are widely distributed in the ocean can secrete diarrheal shellfish toxins. This toxin is transferred and accumulated among organisms at various trophic levels through the food chain. If poisoned shellfish is eaten by mistake, diarrhea, nausea, Vomiting and other poisoning symptoms, the main symptoms of poisoning are diarrhea and vomiting, so it is also called diarrhea shellfish poisoning; and because most of the DSP poisoning incidents are caused ...

AI Technical Summary

Problems solved by technology

[0004] Commonly used detection methods for diarrheal shellfish toxins include mouse biological method, liquid chromatography and enzyme-linked immunosorbent assay. Dry, then dissolve the residue with 1% Tween-60 normal saline, inject mice intraperitoneally, observe the survival situation, and calculate its toxicity. Due to the long time of mouse biological detection, the composition of the toxin cannot be determined, and the quantification is difficult. High positive (usually caused by fatty acids), poor reproducibility, has been rarely used; liquid chromatography is the sample is extracted by methanol, hydrolyzed under alkaline conditions to release esterified diarrhea shellfish toxin, liquid chromatography Separation, tandem mass spectrometry, and quantitative analysis are by far the most widely used and most recognized detection methods; enzyme-linked immunosorbent assay mainly uses the interaction between the antibody of diarrheal shellfish toxin and the antigen in the sample, through Chromogenic reaction is used to detect diarrheal shellfish toxin in the sample. This method is convenient to use and has high detection sensitivity. However, due to the complexity of the detection sample and the large number of matrices, the probability of false positive detection is high.

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