Synthesis And Identification Of The Immunogen Of Cyclosporin A

OBJECTIVECsA is a micromolecular undecapeptide. It is a hapten without immunogenicity and lacks of active group which could connect with carrier protein. The objective of this experiment is to design correct synthetic route, optimize synthesis technology, and synthesis the Immunogen of Cyclosporin A. First, modify its structure to introduce such active functional groups. Then couple with carrier protein or polypeptide forming a whole antigen. Last, synthesis the Immunogen of Cyclosporin A, which is a preparation for manufacturing the kit of fluorescence polar immunologic surveillance.METHODSChoose the photochemistry reaction to modify CsA's structure, and introduce the active functional group, carboxy. The BBa and CsA together can synthesize CsA-BBa intermediate through hydrogen-abstraction and cycloaddition reaction with the help of high voltage mercury lamp as ultraviolet light source and tert-butyl alcohol as reaction solvent. The experiment use orthogonal experiment to optimize and determine optimal condition. Choose PL as protein carrier in the coupling reaction. Dissolve CsA-BBa in DMF and synthesize CsA-BBa-PL complete antigen with PL under the condition of EDCI in water solution.RESULTSTest CsA, BBa and CsA-BBa by melting point, lamella, IR,1H-NMR,13C-NMR and MS, proving successfully synthesizing of CsA-BBa. Detect the complete antigen Cyclosporin A by ELISA, confirming the complete antigen can react with monoclonal antibody Cyclosporin A specifically.CONCLUSIONWe successfully find the way to synthesize the complete antigen CsA and it may be applied in Cyclosporin A assay with FPIA.