| Clenbuerol is a potent respiratory stimulant used in human and animal medicine. This drug is , however , used illegally as a growth promoter in production animals ,because its use leads to carcass which are fat-free and which have an extraordinary development of muscular mass. While much of the Clenbuterol will be excreted or metabolized, there is still a considerable amount retained by the animals especially in the liver, muscles, and retina. As a results , the residues rendered a great danger to consumer and animal ,furthermore ,several food poisoning accidents related to the consumption of illicit β 2 agonist in liver was happened in China and other country. There is a great demand about techniques related to the detection of residue and safety drug which have Clenbuerol positive effect and no lexicological characteristics.According to this, the content in this paper is included as follow : Synthesis and Identification of Clenbuterol -carrier protein conjugate, the Preparation of Monoclonal Antibody against Clenbuterol and Cloning & expression of (32 Adrenergic Receptor Gene of Mouse.For the reason that small moleculars liking Clenbuterol have their immunoreactivity but no immunogenicity, thus they couldn't induce animal to produce antibody until they have been conjugated to macromolecular ,such as bovine serum albumin, human serum albumin. Unfortunately ,little study has been done to make sure that small molecular ,hapten ,has been conjugated with carrier protein. As a result, no effective method has been put forward . In this paper, some work have been done and tried to solve this problem . A conjugate prepared by the direct coupling of diazotized Clenbuterol to bovine serum albumin was identified by IR, UV , SDS-PAGE v FPLC and immunological experiment . Compared with each other, a brief assessment of each analytical method is given. A conclusion was draw that among all the analytical methods, FPLC is the best one.Monoclonal antibody was prepared by classical hybridomas technique . The whole process included the preparation of antigen(enough forimmunization and screen),planning a screening assay, developing the screening assay, immunization the mouse, preparation the sp2/0 cell, plasma cells in spleen ,feeding cell, the fusion of the sp2/0 cell and plasma cells in spleen cell , screening to get positive clone and cloning ,colony growth. After the whole operation , positive clone which could secrete antibody against Clenbuterol was obtained, furthermore the antibody concentration was measured. Monoclonal antibody could use as standard reagents in the residue immunoassay and reagents in the treatment of sample.There are two way to obtain safety drug , on the one hand, when a pharmaceutical screening model has been established , easy-to-catabolism bioactive molecular could be approached . on the other hand, antibody against β sadrenoceptor, which is binding with the receptor just as β 2 adrenergic stimulant, may have the same bioactivity, anti-idiotypic antibodies against anti-Clenbuterol Monoclonal Antibody could Mimics the bioactive ofβ2 adrenergic stimulant. Above all, the basis of these study is that the β 2 adrenergic receptor gene of mouse must have been cloned and the monoclonal antibody against Clenbuterol must been prepared successfully.β2 Adrenergic receptor Gene of Chinese Kunming mouse was amplified by PCR and it was ligated with pUCm-T vector by T4 ligase . The ligation product was then transfered into competent E.coli DH5α . After screening with α-complementarity, positive clone was identified by PCR ,restrict enzyme characterization and DNA sequencing . Furthermore ,the gene was blast in NCBI . A conclusion was draw that β2 adrenergic receptor gene of mouse had been cloned. Then,β2 Adrenergic receptor gene was expressed in E.Coli and fusion protein was detection by SDS-PAGE .All in all , The meaning all work in this paper is that providing technique associated with residue detection , lay a basis for the screening safety drug .
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