Preparation Of Monoclonal Antibody Against Salbutamol And Establishment Of Immunoassay Method For Salbutamol

Salbutamol(Sal),also called tetrahydro-isowuinolin,is a kind of β₂receptor agonist which can selective agitate the β₂- receptor of TSM .it is used to treat BA clinically .The stimulant of β₂ just as Sal is the Avian of growth promotion agt(such as cattle, caprine, fowl and swine and so on),many researchers have extensively studied it's effective- ness,and it was used to breed livestock and fowl generally. But the stimulant of β₂ is often remained in creature's organs , even enter human's body by food chain and seriously jeopar- dizing human's health .with many countries,such as Spain, Holland, France and China, were empoisoned ,so many states make laws to forbid using stimulant of β₂ as additives in forage one by another. Many states paied close attention to Clenbuterol for it can redistribute nutrition and was used extensively in cultivation , now many states have already founded a series of quick and effective technology to monitor the content of Clenbuterol ,the technology prevent Clenbuterol to be abused .Sal have already become the substitute of Clenbuterol for the technology of detect sal is relative hysteresis, even it have the same effection with Clenbuterol.It is a stringent task to found a simple, convenient, effective and quick technology to detect Sal be abused.Conjugating Sal to bovine serum albumin (BSA)and ovi albumin (OVA) by succinic anhydride and carbodiimide transformed Sal,an half antigen,into complete antigen.The productions are Sal-HS-BSA and Sal-HS-OVA. Adequately dialysis is required to purify the product.Then confirm that Sal has been conjugated to BSA and OVA. To obtain purified special mice anti-Sal,four Blab/c mice were immunized with BSA-Sal.Antibody titers were determined using indirect ELISA with OVA-Sal in coating the micro-wells.we can conjugate cell when the potency reach over 1:10000.Cell fusion was Proceeded in a 50-ml centrifuge tube.Mingle spleen cells and myeloma cells (5:l)into the centrifuge tube,and then instill PEG-1000 scrupulously at a certain dose.Put the mixed cells into the cell culture wells that had been paved with feeder cells,and selected-culture with HAT and HT.Avoiding contamination is crucial to the cell fusion.Inspect the cells next day.Small clones can be seen in the sixth day.