The Design And Initial Application Of Test Papers For The Rapid Detection Of Blood Concentration Of Cyclosporin A

Partâ… Synthesis of cyclosporine A complete antigenObjectives To prepare the cyclosporin A complete antigen,and inspect its correlative speciality.Methods We connect a acticve chemical groups to cyclosporin A, couple cyclosporin A which is modified with carrier protein(bovine serum albumin) to synthesize cyclosporin A complete antigen, then purify it by dialysis. UV scan and SDS-PAGE is used to confirm whether cyclosporin A have connected to the carrier protein, and compute the coupling rate.Results We obtain cyclosporin A complete antigen. Both UV scan and SDS-PAGE reveal cyclosporin A have connected to the carrier protein,and the coupling rate is 18.3:1.Conclusion Cyclosporine prepration of A complete antigen is obtained. Partâ…¡preparation of cyclosporin A polyclonal antibodyObjectives To prepare cyclosporin A polyclonal antibody,and inspect its correlative speciality.Methods We use Freund's adjuvant to emulsify the antigen, immune animal some times,and obtain blood from femoral artery. Antibody titer is tested by indirect-ELISA, and specificity of antibody is evaluate by 50% inhibiting concentration ( IC50 ) . Salting out is used to purify the antibody.The content of protein in the products is determined by Coomassie Brilliant Blue method.Results We obtain the antiserum after immune five times. Antibody titer is 1:16000,and it is specific against some clinical drugs such as Lansoprazole, cefradine, ambroxol, glibenclamide. The content of protein after purifying is 1.367mg/ml.Conclusion The cyclosporin A polyclonal antibody is obtained. Partâ…¢preparation of test papers for rapid detection of blood concentrateon of cyclosporin AObjectives To prepare test papers for rapid detection of blood concentrateon of cyclosporin A, to evaluate its specificity, and to detect clinical samples using test papers for rapid detection of blood concentrateon of cyclosporin A.Methods Cyclosporin A polyclonal antibody was marked by colloidal gold to mark,and add it on the cellulose membranes, then assemble into a test papers.We choose appropriate the cellulose membranes, optimize technological parameter,evaluate some indexes such as sensitivity,persion,detection range, limit of quantification,stability and so on. We detect clinical samples and analyse its effectiveness by comparing the results to TDX.Results The detection range of test papers is 5-600ng/ml. Formula of standard curves is A=-0.1903C+143.69. There is no obvious cross reactivities for Lansoprazole, cefradine, ambroxol, glibenclamide. The difference of blood concentration detected between test papers and TDX is not distinct.Conclusion Test papers for rapid detection of blood concentrateon of cyclosporin A has been preliminary developed,but it may also need some further research.