Hybridoma cell strain and anti-hydrochloric acid clenbuterol monoclonal antibody prepared from hybridoma cell strain

A technology of clenbuterol hydrochloride and hybridoma cell lines, which is applied in the field of immunochemistry, can solve the problems that antibodies cannot meet the needs of detection, low specificity, low specificity of monoclonal antibodies, etc., and achieve rapid and sensitive detection Effect

Active Publication Date: 2011-01-12
JIANGSU HUACHUANG MEDICINE RES & DEV PLATFORM MANAGEMENT CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

This patented technology allows us to produce very small amounts (<1 milligram) of an active ingredient called clonidone that blocks nerve impulse transmission at certain sites on brain cells. These tiny particles could potentially help treat neurologically related conditions such as Parkinson’s disease or Alzheimer's diseases.

Problems solved by technology

This patents discusses different ways how drugs likeclenevent® or other medications are being given up overweight due to their side effects when taken regularly without adequate monitoring during therapy sessions. These agents have been found to cause harmful deposits within the patient's gastrointestinal tract leading to various complications including dysfunctional bradycardics and pulmonephritis syndrome. To address these issues, there is an increasing interest among manufacturers who want to develop effective techniques for identifying residue formation from animal sources while minimizing any potential negative impact upon humans.

Method used

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  • Hybridoma cell strain and anti-hydrochloric acid clenbuterol monoclonal antibody prepared from hybridoma cell strain
  • Hybridoma cell strain and anti-hydrochloric acid clenbuterol monoclonal antibody prepared from hybridoma cell strain
  • Hybridoma cell strain and anti-hydrochloric acid clenbuterol monoclonal antibody prepared from hybridoma cell strain

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Experimental program
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Embodiment 1

[0029] Reagent and material preparation

[0030] Clenbuterol hydrochloride standard substance (Beijing Shanglifang United Chemical Technology Research Institute, BW8001), concentrated hydrochloric acid (Shanghai Pilot Chemical Corporation), sodium carbonate (Sigma, 22353-0), sodium bicarbonate (Sigma, S6297), Sodium nitrite (Shanghai Yanchen Chemical Industry Co., Ltd.), ammonium sulfamate (Sinopharm Chemical Reagent Co., Ltd.), bovine serum albumin (BSA) (Amresco, 0332), human serum albumin (HSA) (Sigma, A9511 ), ovalbumin (OVA) (Sigma, A5503), Freund's complete adjuvant (Sigma, F5881), Freund's incomplete adjuvant (Sigma, F5506), tetramethylbenzidine (TMB) (Amresco, 0759) , HAT (Sigma, H0262) and HT (Sigma, H0137), sodium chloride (Amresco, 0241), potassium chloride (Amresco, 0395), potassium dihydrogen phosphate (Sigma, P9791), disodium hydrogen phosphate (Sigma, 71639), goat anti-mouse IgG-HRP (Jackson, 115-035-044), dimethyl sulfoxide (DMSO) (Applichem, 0231), polyethyle...

Embodiment 2

[0112] Embodiment two drug cross reaction test

[0113] According to the indirect competition ELISA method established in Example 1, the monoclonal antibody 6G11' of clenbuterol hydrochloride and clenbuterol hydrochloride hapten structural analogues such as salbutamol and ractopamine were respectively subjected to competition tests. Dilute these standards into different concentrations for indirect competition ELISA experiments, draw inhibition curves, and calculate ICs of competitors 50 value and cross-reactivity rate, the highest cross-reactivity rate was 2.7%. The monoclonal antibody of clenbuterol hydrochloride reported in the literature has a cross-reactivity rate of 50.04% with salbutamol, and a cross-reaction rate of 75% with ractopamine, while the anti-clenbuterol monoclonal antibody prepared by the present invention It only reacts with clenbuterol hydrochloride, therefore, the anti-clenbuterol hydrochloride monoclonal antibody prepared in the present invention has hig...

Embodiment 3

[0116] Example 3 Application of Monoclonal Antibody 6G11' in the ELISA Method for Detecting Clenbuterol Hydrochloride Residues

[0117] The monoclonal antibody 6G11' can be applied to the ELISA method for detecting clenbuterol hydrochloride residues, for example, it can be used for the detection of clenbuterol hydrochloride residues in bull's eye, urine, and muscle. Taking pork (purchased in the market) as an example, the main steps for detecting clenbuterol hydrochloride residues in pork are as follows:

[0118] 1. Sample pretreatment: adopt general method for processing.

[0119] 2. The detection principle of the kit in the present invention is the indirect competition ELISA method, the original OVA-CL coated on the microporous plate is coated, the clenbuterol hydrochloride standard or sample diluted in series is added, and the anti-hydrochloric acid gram Renbuterol monoclonal antibody 6G11', the residual clenbuterol hydrochloride in the sample and the coated antigen compet...

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Abstract

The invention provides a hybridoma cell strain and an anti-hydrochloric acid clenbuterol monoclonal antibody prepared from the hybridoma cell strain and belongs to the technical field of immunochemistry. The hybridoma cell strain 6G11 generates the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11'. The invention also provides a method for preparing the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11', which comprises the following steps of: preparing an immunogen and a coating antigen; animal immunization; selecting immune rat serum; preparing hybridoma cells; and collecting and purifying the anti-hydrochloric acid clenbuterol monoclonal antibody 6G11'. The anti-hydrochloric acid clenbuterol monoclonal antibody with strong specificity through the rat hybridoma cell strain 6G11, has low crossing reaction rate with the salbutamol and ractopamine, can be massively produced, and can be used for preparing immunology detecting agents, such as an immunoassay kit for detecting clenbuterol hydrochloride, a colloidal gold test strip and the like so as to fast and sensitively detect clenbuterol hydrochloride medicament residue in urine, muscles and internal organs.

Description

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Claims

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Application Information

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Owner JIANGSU HUACHUANG MEDICINE RES & DEV PLATFORM MANAGEMENT CO LTD
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