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Preparation method of horse radish peroxidase labeled leuco-malachite green immunogen

A technology of horseradish peroxidase and colorless malachite green, which can be used in measuring devices, instruments, scientific instruments, etc., can solve the problems of unfavorable colorless malachite green residue detection, large drop in antibody titer, etc. The marking effect is ideal, which is conducive to popularization and application, and the effect of improving accuracy and sensitivity

Inactive Publication Date: 2014-04-16
SHANGHAI OCEAN UNIV
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AI Technical Summary

Problems solved by technology

[0002] At present, in the detection of colorless malachite green, the common labeling method is to label the antibody, but the labeling of the antibody has the disadvantage that the antibody titer drops greatly after labeling, which is not conducive to the detection of colorless malachite green residues

Method used

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Embodiment Construction

[0013] The preparation method of the horseradish peroxidase-labeled leuco-malachite green immunogen of the present invention will be further described in detail below.

[0014] Specific steps are as follows:

[0015] (1) Weigh 5mg of horseradish peroxidase into a 10mL centrifuge tube, add 1mL of double distilled water to dissolve, then slowly add 0.2mL of freshly prepared 0.1mol / L sodium periodate solution while shaking slowly, at room temperature React in the dark for 20 minutes, put the reacted liquid into a dialysis bag, then put it into 1mmol / L pH4.4 sodium acetate buffer, dialyze in the dark at 4°C for 24 hours, change the liquid 4 times during the period, and then collect the dialysis bag Liquid, get the horseradish peroxidase solution that dialyzes;

[0016] (2) First add 0.2mol / L pH9.5 carbonate buffer solution to 1mL colorless malachite green immunogen solution, adjust the pH to 9.1, and then dialyze the horseradish peroxidase in step (1) Adjust the pH of the soluti...

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Abstract

A preparation method of a horse radish peroxidase labeled leuco-malachite green immunogen is characterized in that: the synthesized leuco-malachite green immunogen is labeled by horse radish peroxidase by using a sodium periodate method. The labeling method comprises the following steps: the carbohydrate chain of the horse radish peroxidase is oxidized by the sodium periodate at the room temperature and low pH value, the obtained hydroformylated horse radish peroxidase is linked with the amino group of the leuco-malachite green immunogen and then the link body is reduced by sodium borohydride to form the stable horse radish peroxidase labeled leuco-malachite green immunogen; the horse radish peroxidase labeled leuco-malachite green immunogen precipitates by adding 50% saturated ammonium sulfate, the precipitate of the precipitate horse radish peroxidase labeled leuco-malachite green immunogen is dissolved by a phosphate buffer, and finally the solution is dialyzed to obtain the horse radish peroxidase labeled leuco-malachite green immunogen. The horse radish peroxidase labeled leuco-malachite green immunogen capable of detecting the residue of the leuco-malachite green is successfully synthesized, and moreover the invention establishes the foundation for development of the leuco-malachite green medicine residue enzyme-linked immunosorbent assay, and also facilitates popularization and application.

Description

technical field [0001] The invention relates to a preparation method of horseradish peroxidase-labeled immunogen, in particular to a preparation method of horseradish peroxidase-labeled colorless malachite green immunogen. Background technique [0002] At present, in the detection of colorless malachite green, the common labeling method is to label the antibody, but the labeling of the antibody has the disadvantage that the antibody titer drops greatly after labeling, which is not conducive to the detection of colorless malachite green residues. Contents of the invention [0003] The purpose of the present invention is to provide an improved preparation method of horseradish peroxidase-labeled leuco-malachite green immunogen, which can not only effectively produce enzyme-labeled leuco-malachite green immunogen for detection of colorless malachite green residue Malachite green immunogen, and easy to operate, ideal labeling effect. [0004] The object of the present inventi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535
CPCG01N33/535
Inventor 杨先乐梁楠柯江波胡利静刘腾飞
Owner SHANGHAI OCEAN UNIV
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