Development Of Enzyme-Linked Immunosorbent Assay For Ractopamine

β- Adrenergic agonist have been used as bronchiodilators in human medicine for over 30 years.Unfortunately, these agents have also been used illegally to enhance the growth rates of livestocks treated with such drugs by promoting the repartitioning of fat into mucle.There illegal use has led to toxic effects after human consumption of meat products.Ractopamine is a phenethanolamine member of the family of β - Adrenergic agonist. Except a few country such as American, a lot of country(such as Eu and china) in the world have all banned the use of growth promoters.When the β -agonist such as clenbuterol was banned the use of β - Adrenergic agonists as growth promoters in livestock in our country, the illegal use of ractopamine in livestock was carried out by illegal enterprise and manufacturer, because of potential for illegal ractopamine use,there is a need for rapid detection methods.The official regulatory determinative method of ractopamine analysis such as LC,HPLC,LC/MS,GC/MS requires multiple extractions,so the rapid screening method should be developed.Now,there is only USA to develop the ELISA for ractopamine.In our study,multi-anhydride combined with mixed anhydride was used to couple the ractopamine with carrier proten(KLH and BSA). Antibody generated from ractopamine-hemiglutarate-KLH was use to develop ELISA for ractopamine. Ractopamine-hemiglutarate-BSA was used to coating antigen to detect the antibody titers.The conjugation and conjugation ratio was characterized by NMR and UV spectrametry, The different conjugation ratio artificial antigen was synthesized with changing the carrier protein concentration in solvent.The antibody showed good specificity with an IC50 of 10 ng/ml toward ractopamine.The antibody didn't show cross reactivity towards clenbuterol,salbutamol and terbutaline.The indirect competitive ELISA curve and the direct competitive ELISA curve was developed . The limit of detection of the indirect competitive ELISA curve was approximately 0.9ppb, it's recovery ranged between 63.5% and 114.%.The CV% ranged from 3.1% to 16.7%. The limit of detection of the competitive ELISA curve was approximately lppb, it's recovery ranged between 89.4% and 144.1%. The CV% ranged from 3.05% to 14.36%.The quality control of the ELISA was evaluated in the chapter 5.In this paper,the influence facture of synthesis of artificial antigen and specificity of antibody was discussed. The imagination about relevance between the animal type of build parameter and immune responsivity was first bring up.