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Preparation method of metandienone polyclonal antibody immuno-affinity column and application thereof

A technology of polyclonal antibody and dehydromethyltestosterone, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as interference, lack of selectivity, and cumbersome steps

Inactive Publication Date: 2011-09-14
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These instrument detection methods have high requirements on sample purity and require certain pretreatment, while traditional sample pretreatment techniques, such as liquid-liquid extraction (LLE), solid phase extraction (SPE), solid phase microextraction (SPME), etc. , the steps are cumbersome, lack of selectivity, easily interfered by other components, and low accuracy
But there is no report on the preparation and use of DMT polyclonal antibody immunoaffinity column

Method used

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  • Preparation method of metandienone polyclonal antibody immuno-affinity column and application thereof
  • Preparation method of metandienone polyclonal antibody immuno-affinity column and application thereof
  • Preparation method of metandienone polyclonal antibody immuno-affinity column and application thereof

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Embodiment 1

[0022] Example 1 Preparation of DMT polyclonal antibody immunoaffinity column

[0023] (1) Coupling

[0024] Place the immunized animals and the purified DMT polyclonal antibody in 0.1 mol / L NaHCO containing 0.5 mol / L NaCl at pH 8.3 3 Dialyze in coupling buffer overnight, and dilute to 0.5 mg / mL before use;

[0025] Weigh CNBr-Sepharose 4B dry gel, fully swell in 5-10 mL of 1 mmol / L HCl solution to obtain a certain volume of wet gel. Quickly wash twice with coupling buffer more than 10 times the volume of the wet gel, and centrifuge at 4000 r / min for 1 min;

[0026] Dilute the washed wet gel to 2 mL, quickly add 0.5 mg / mL antibody solution, make the mass ratio of CNBr-Sepharose 4B dry gel to antibody 100:1, shake and react at room temperature for more than 2.5 h;

[0027] After the coupling reaction, wash with coupling buffer that is more than 10 times the volume of the coupling gel to remove unconjugated antibodies, collect the washing liquid, and use formula C protein (m...

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Abstract

The invention relates to a preparation method of a metandienone polyclonal antibody immuno-affinity column and an application thereof, and belongs to the technical fields of immuno-affinity chromatography and veterinary drug residue detection. A preparation method and an application of a metandienone polyclonal antibody immuno-affinity column are disclosed. A immuno-affinity column with specific adsorption to metandienone is prepared by purifying a polyclonal antibody obtained from an immunized animal, coupling the antibody with CNBr-Sepharose4B to obtain an immuno-affinity adsorbent, and filling a solid-phase extraction tube with the adsorbent. The immuno-affinity column prepared by the invention can specifically bind to metandienone with a maximal binding capacity of 160 ng DMT; when an eluate of an 80% methanol-aqueous solution is used, the average recovery rate of standard addition is 97.87%; after repeated use for 3 times, the recovery rate is not less than 90%. The invention is applicable to the rapid detection of metandienone.

Description

technical field [0001] The invention belongs to the technical field of immunoaffinity chromatography and detection of veterinary drug residues, and relates to a preparation method and an application of a dehydromethyltestosterone polyclonal antibody immunoaffinity column. Background technique [0002] Dehydromethyltestosterone (1-dehydro-17-methyltestosterone, DMT) is a protein anabolic androgenic steroid hormone. Because of its strong effects of promoting protein synthesis, increasing appetite, and improving feed conversion rate, it is often illegally used in the field of animal husbandry. After DMT is absorbed by the human body, it will interfere with the body's own hormone balance and cause serious toxic and side effects. [0003] At present, the commonly used detection methods for DMT are mostly instrumental detection methods, such as high performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and gas chromatography-mass spectrometry ...

Claims

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Application Information

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IPC IPC(8): G01N30/50G01N30/56G01N30/06
Inventor 王云徐燕董英张勋
Owner JIANGSU UNIV