Affinity chromatography-enzyme linked immunity test method for ractopamine and dedicated kit

A ractopamine and enzyme-linked immunosorbent assay technology, which is applied in the field of immunoassays, can solve the problems of inability to adapt to fast, simple, inexpensive, difficult mass detection, and time-consuming instrumental analysis techniques.

Inactive Publication Date: 2005-10-26
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These instrumental analysis techniques are generally time-consuming, expensive, difficult to detect in large quantitie

Method used

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  • Affinity chromatography-enzyme linked immunity test method for ractopamine and dedicated kit
  • Affinity chromatography-enzyme linked immunity test method for ractopamine and dedicated kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] 1. Sample pretreatment:

[0087] Weigh 10g of pork products (including liver, lung, heart, kidney, muscle), add 50ml 0.1mol / L HCl for tissue homogenization, freeze and thaw twice, centrifuge to get the supernatant, adjust the pH value to 11 with 10mol / L NaOH, Then add 30ml of isobutanol, shake for 30min, let it stand for 4h, quantitatively absorb isobutanol, evaporate to dryness in a water bath, then add 1ml of phosphate solution (PBS, 0.01mol / L, pH7.4) to dissolve the eluate, and freeze it for testing ;

[0088] Pig urine samples were centrifuged to remove impurities, and the supernatant was taken for testing.

[0089] 2. Preparation of enzyme-labeled antigen: mix activated horseradish peroxidase with purified RCT, perform labeling reaction in carbonic acid buffer system, and then add sodium borohydride to reduce unbound sites;

[0090] 3. Preparation of RCT antibody

[0091] Coupling RCT with bovine serum albumin BSA to prepare antigen; immunization dose 1mg / ml BSA...

Embodiment 2

[0107] The composition of the RCT detection kit:

[0108] The kit includes: ELISA plate coated with monoclonal RCT antibody; A reagent: standard RCT reagent; B reagent: diluent; C reagent: enzyme-labeled antigen reagent; D reagent: enzyme-labeled antigen diluent; E reagent: containing Tween -20 phosphate PBS solid; F reagent: o-phenylenediamine reagent; G reagent: sodium acetate-citric acid buffer; H reagent: 30% H 2 o 2 ); I reagent: sulfuric acid solution; J reagent: glycine solution; K reagent: sepharose-4B gel coupled with monoclonal RCT antibody.

Embodiment 3

[0110] The composition of the RCT detection kit:

[0111] The kit includes: ELISA plate coated with polyclonal RCT antibody; A reagent: standard RCT reagent; B reagent: diluent; C reagent: enzyme-labeled antigen reagent; D reagent: enzyme-labeled antigen diluent; Phosphate PBS solid at temperature -20; F reagent: o-phenylenediamine reagent; G reagent: sodium acetate-citric acid buffer; H reagent: 30% H 2 o 2 ); Reagent I: sulfuric acid solution; Reagent J: glycine solution; Reagent K: sepharose-4B gel coupled with polyclonal RCT antibody.

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Abstract

This invention uses immunity compatible chromatography post and enzyme linked immunosorbent assay to detect the remain RactopamineHy-drochloridie in food. It belongs to immunology and food safety detect technology. The method is: prepare muti-clone antibody by immunity antigen synthesis, antigen peridium, animal immunity and antigen depuration. Then put the antigen on to agarose gel to prepare immunity compatible chromatography post. You can enrich the RactopamineHy-drochloridie in the sample by compatible post and gather the eluent of the compatible post, then use linked immunosorbent assay to determine the consistency of the RactopamineHy-drochloridie.

Description

technical field [0001] The invention belongs to the technical field of immune analysis. Specifically, it relates to a ractopamine (RCT) affinity chromatography-ELISA detection method and a special detection kit thereof. Background technique [0002] Ractopamine is one of the members of β2-adrenergic agonist (BAA), and it belongs to catecholamines (adrenaline and norepinephrine BAA) substances like "leptbuterol". As early as the late 1950s, it was discovered that catecholamines (epinephrine and norepinephrine) could stimulate fatty tissue to release fatty acids and increase protein deposition, but it was not until the early 1970s that catecholamines and their analogs improved the body's nutritional allocation The ability to form and carcass was first recognized by the American Cyamide Research Department. Then it is used on a large scale in livestock and poultry production to increase lean meat percentage, reduce fat deposition, and increase milk production. [0003] The p...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 黄昆仑许文涛邓爱科罗云波
Owner CHINA AGRI UNIV
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