Tetracycline antibiotics immunoaffinity column and preparation method and application thereof

A technology of tetracyclines and antibiotics, applied in chemical instruments and methods, separation methods, and other chemical processes, can solve problems such as high-concentration drug residues, superinfection, and impact on consumers' health, and achieve large binding capacity and specificity. The effect of high specificity and improving antibody binding ability

Inactive Publication Date: 2018-09-21
SHANDONG MEIZHENG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Tetracycline antibiotics (Tetracyclines) are a class of broad-spectrum antibiotics produced by actinomycetes, including chlortetracycline (chlortetracycline), oxytetracycline (oxytetracycline), tetracycline (tetracycline) and semi-synthetic derivatives methoxytetracycline, strong Aminocycline, dimethylaminotetracycline, etc., all of which contain naphthacene basic skeleton; among them, tetracycline, oxytetracycline, chlortetracycline and doxycycline have excellent antibacterial properties, stable drug properties and low prices. It is widely used in the treatment and prevention of diseases and growth promotion in animal husbandry production, but excessive use of such drugs will lead to high concentration of drug residues in animal products, directly affecting the health of consumers, adverse reactions of tetracycline antibiotics include gastrointestinal reactions , superinfection (flora alternation syndrome), effects on bone and tooth growth, liver damage, vitamin deficiency and nephrotoxicity, in addition, tetracycline antibiotics can occasionally cause allergic reactions such as drug fever and rash

Method used

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  • Tetracycline antibiotics immunoaffinity column and preparation method and application thereof
  • Tetracycline antibiotics immunoaffinity column and preparation method and application thereof
  • Tetracycline antibiotics immunoaffinity column and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The main structure of the tetracycline antibiotic immunoaffinity column involved in this embodiment includes a column body 1, a cover plate 2, an injection port 3, a first sieve plate 4, a filler 5, a second sieve plate 6 and a sample outlet 7; The top of the cylinder 1 with a tubular structure is provided with a cover plate 2 with a circular sheet structure, and the center of the cover plate 2 is provided with a sample inlet 3 with a circular tubular structure. The inner upper space of the cylinder 1 is a cavity, and the cylinder 1 The first sieve plate 4 with a circular sheet structure with a sieve hole is arranged in the middle of the interior of the cylinder, the inner lower space of the column 1 is filled with fillers 5, and the bottom of the column 1 is provided with a circular sheet with a sieve hole. The second sieve plate 6 of the structure, the second sieve plate 6 is provided with a sample outlet 7 of a conical tubular structure with a wide top and a narrow bo...

Embodiment 2

[0025] The specific process of the tetracycline antibiotics immunoaffinity column preparation method involved in this embodiment includes five steps of activation, coupling, connection, cross-linking and column packing:

[0026] (1) Activation: fully rinse the pre-swelled agarose gel sepharose4B with a concentration of 2% with distilled water, wash away the remaining ethanol in the agarose gel sepharose4B, the volume ratio of distilled water and agarose gel sepharose4B is 20:1, use Funnel filters liquid, obtains wet gel, takes by weighing 5 grams of wet gel and 7.5 milliliters of molar masses and is the NaOH (sodium hydroxide) aqueous solution of 0.8M, 2 milliliters of mass percentage concentrations and is 30% epichlorohydrin aqueous solution and 5 mL of NaBH at a concentration of 2 mg / mL 4 (Sodium borohydride) aqueous solution is carried out shaking table reaction under the condition of 25 ℃ for 8 hours at a temperature, after the reaction finishes, wash the wet gel with 50 m...

Embodiment 3

[0032] The tetracycline antibiotic immunoaffinity column that the present embodiment involves is purified and the concrete technological process of the tetracycline antibiotic in the enrichment feed is: first, in the No. 50 milliliter centrifuge tube that is placed with 5.0g homogeneous chicken, add 25 Milliliter of 70% methanol aqueous solution, vortexed for 2 minutes, then centrifuged at 12000rpm for 10 minutes, transferred the supernatant to No. 2 50 ml stoppered centrifuge tube, and then added Volume is 6 times of the supernatant volume concentration and is that the PBS of 10mM dilutes the supernatant, obtains the tetracycline antibiotics feed solution, finally, regulates the air pump pressure of the air control operation frame, makes the tetracycline antibiotics feed solution pass through the syringe with 2 The flow rate of -3 drops / second flows through the tetracycline antibiotic immunoaffinity column, and after the tetracycline antibiotic feed solution all flows out of t...

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Abstract

The invention belongs to the technical field of food safety inspection, and particularly relates to a tetracycline antibiotics immunoaffinity column and a preparation method and application thereof. Sepharose gel activated through an epichlorohydrin activation method is taken as a carrier, gene recombinant protein G is coupled to the carrier to obtain protein G-sepharose, a tetracycline antibiotics antibody is connected to the protein G-sepharose to obtain antibody-protein G-sepharose, a cross-linking agent is used for crosslinking, antibody-protein G-sepharose fillers are obtained, and finally, the immunoaffinity column is filled with the antibody-protein G-sepharose fillers. The specific binding feature of the gene recombinant protein G and tetracycline antibiotics is fully utilized, Fabfragments of the tetracycline antibiotics are exposed to the outside, and antibody fixation capacity of the gene recombinant protein G, capture capacity of the tetracycline antibiotics and purification efficiency of the purification efficiency are greatly improved.

Description

Technical field: [0001] The invention belongs to the technical field of food safety detection, and in particular relates to a tetracycline antibiotic immunoaffinity column and its preparation method and application. 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] Tetracycline antibiotics (Tetracyclines) are a class of broad-spectrum antibiotics produced by actinomycetes, including chlortetracycline (chlortetracycline), oxytetracycline (oxytetracycline), tetracycline (tetracycline) and semi-synthetic derivatives methoxytetracycline, strong Aminocycline, dimethylaminotetracycline, etc., all of which contain a naphthacene basic skeleton; among them, tetracycline, oxytetracycline, chlortetracycline and doxycycline have excellent antibacterial pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/20B01D15/22B01D15/38B01J20/281G01N33/531
CPCB01D15/3823B01D15/206B01D15/22B01J20/281G01N33/531
Inventor 柳家鹏王涛于蓉龚利新
Owner SHANDONG MEIZHENG BIOTECHNOLOGY CO LTD
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