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Phage antibody library and application thereof in content determination of clenbuterol hydrochloride

A phage antibody library and clenbuterol hydrochloride technology, applied in the field of molecular biology, can solve problems such as difficult to achieve accurate dosage of clenbuterol hydrochloride, low accuracy of clenbuterol hydrochloride, etc., to achieve easy implementation, The effect of fast detection speed and low detection cost

Active Publication Date: 2013-10-23
WEIFANG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the immunoassay method has the advantages of rapidity, strong specificity and less sample requirement, the accuracy of the disclosed several antibodies for detecting clenbuterol hydrochloride is relatively low, so that the consumption of clenbuterol hydrochloride is still relatively low. Difficult to achieve accurate use

Method used

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  • Phage antibody library and application thereof in content determination of clenbuterol hydrochloride
  • Phage antibody library and application thereof in content determination of clenbuterol hydrochloride
  • Phage antibody library and application thereof in content determination of clenbuterol hydrochloride

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Embodiment

[0045] 1. Construction of phage antibody library

[0046] Material:

[0047] Clenbuterol hydrochloride and bovine serum albumin conjugate (CLEN-BSA) were purchased from Hangzhou Nankai Rixin Biotechnology Co., Ltd.; free clenbuterol hydrochloride was purchased from China Institute for the Control of Pharmaceutical and Biological Products; total RNA extraction kit was purchased from Beijing Biotech Biotechnology Co., Ltd.; Escherichia coli DH5α was purchased from TaKaRa Biotechnology Co., Ltd.; Escherichia coli TG-1 was purchased from Amersham Bioscience (Tokyo, Japan); restriction endonucleases and polymerases used for gene cloning were purchased from TaKaRa Bio Inc. All other reagents are of domestic analytical grade unless otherwise specified.

[0048] Primer Design:

[0049] In order to amplify the variable domain of antibody heavy chain, we designed and synthesized the VH1-15 forward primer, and added SfiI / NcoI restriction site on the 5' side of the primer.

[0050] MV...

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Abstract

The invention provides a phage antibody library and application thereof in content determination of clenbuterol hydrochloride. A gene VL of an antibody light chain in a variable field is assembled between the enzyme cutting sites of Sall and NotI of phage plasmid pIT2, a gene VH construction of an antibody heavy chain in the variable field is assembled between the enzyme cutting sites of SfiI and XhoI, and the gene of the antibody is obtained after bovine serum albumin and conjugate of clenbuterol hydrochloride are used for immunization of mice. The antibody is the affinity and concentration of single chain antibody library, the bovine serum albumin and the conjugate of the clenbuterol hydrochloride, monoclonal antibody of the clenbuterol hydrochloride is developed to apply to test of residual quantity of the clenbuterol hydrochloride. By using the antibody library, high affinity anti-clenbuterol hydrochloride and chemical substance antibodies with similar structure of the clenbuterol hydrochloride can be obtained directly, and the development and test of the clenbuterol hydrochloride and the chemical substance antibodies with similar structure of the clenbuterol hydrochloride have wide application prospect.

Description

technical field [0001] The invention relates to molecular biology, and is a phage antibody library and its application in the determination of clenbuterol hydrochloride content. Background technique [0002] Clenbuterol hydrochloride (CLEN) is a β2-receptor agonist, belonging to the class of adrenomimetic drugs, with a molecular weight of 313.7. It can improve the metabolic pathways in animals and promote the synthesis of proteins in muscles, especially skeletal muscles. Inhibit the synthesis of fat, thereby accelerating the growth rate, for this reason, many countries are used as feed additives. However, in further research, it was found that excessive intake of clenbuterol can cause huge adverse reactions. In severe cases, symptoms such as acute poisoning, hyperthyroidism, and arrhythmia occur. Therefore, those skilled in the art provide detection methods of liquid chromatography and gas chromatography for controlling the consumption of clenbuterol hydrochloride. Howeve...

Claims

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Application Information

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IPC IPC(8): C40B40/02C40B50/06C12N15/13C12N15/70G01N33/53
Inventor 董金华董益阳
Owner WEIFANG UNIV OF SCI & TECH
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