Development Of The Rapid Immunoassay Strip For The Detection Of Salbutamol

Salbutamol(SAL), is a kind of β-receptor agonist which can selectiveagitate the β-receptor of tracheal smooth muscle.It is used to treat bronchial asthmaclinically.The stimulant of β just as SAL is the Avian of growth promotion agt(suchas cattle, caprine, fowl and swine and so on), and it was used to breed livestock andfowl generally. But the stimulant of β is often remained in creature’s organs，evenenter human’s body by food chain and seriously jeopardizing human’s health.withmany countries were empoisoned, so many states make laws to forbid usingstimulant of β as additives in for-age one by another. Many states paied closeattention to Clenbuterol for it was used extensively in cultivation，now many stateshave already founded aseries of quick and effective technology to monitor the contentof Clenbuterol, SAL have already become the substitute of Clenbuterol for thetechnology of detect SAL is relative hysteresis, even it have the same effection withClenbuterol. It is a stringent task to found a simple, convenient, effective and quicktechnology to detect SAL be abused.In this study, Based on the analysis of molecular structure and immunogenicityof Salbutamol, the technology of monoclonal antibodies production was applied toassemble rapid ELISA kit and strip for SAL residual detection. The main contents andresults of this study were as follows:1. Synthesis and identification of the artificial antigen for SalbutamolSalbutamol was coupled with carrier protein bovine serum abumin (BSA) andovalbumin (OVA) to prepare immunizing antigen BSA-SAL and coating antigenOVA-SAL by the method of mixed anhydride method, Both UV scanning spectrumand SDS-PAGE were used to identify SAL artificial antigen, The result of UVspectrum show that the largest artificial antigen absorption peak migration occurred,SDS-PAGE results show that the BSA’s swimming faster than BSA-SAL, the BSA-SAL molecular weight greater than BSA,This further shows that SAL BSA has beencoupled with success.2. Preparation of monoclonal antibodies and immunological characterization ofSalbutamolOne BALB/C mouse was chosen from four Balb/C mice immunized withBSA-SAL for cell fusion by the identification of Serum testing, The hybridoma lines that fused with NS0myeloma cells and The lymphocytes from Balb/c mouse spleenroutinely by using monoclonal antibody hybridoma technology. One sensitivity,specificity of hybridoma cell lines of4D3E7were screened by using detection,screening and clone, the McAb of4D3E7showed good sensitivity with an IC50of1.35μg/L.9.25%cross-reactivity to Clenbuterol and little or no cress-reactivity toother compounds. SAL McAb obtained can be used to establish immunoassay of SALresidues.3. Development of The Rapid Immunoassay Strip for the Detection of SalbutamolApplication of colloidal gold immunochromatographic test principle (GICA),rapid detection of immuno-gold labeling strips (SAL-Strip) and FITCimmunofluorescence test strips were assembled. Can be used for residues ofsalbutamol in urine test strip can be completed within8-10minutes. Both productsare rapid, sensitive, specific, simple, and can be widely used for the qualitative andsemi-quantitative and quantitative detection of salbutamol residues...