Cyclosporin A Immunoreactive Antigen Synthesis And Testing Of Monoclonal Antibody Research

As a potent immunosuppressant, Cyclosporine A (CsA) has been widely used to prevent rejection of transplanted organs and treat some autoimmune diseases. Due to its special pharmacokinetics and pharmacodynamics, it is very important of monitoring its drug concentration in blood. Among all the methods, immunolabelling technique owning to outstanding feature has been extensively applied. Since this method is based on the affinity reaction between antigen and antibody, so that the key point of success is to obtain specificity antibody. CsA is hapten and without immunogenicity, therefore at first it must be coupled with carrier protein or polypeptide.CsA is a cyclic undecapeptide whose amino acid side chains lack functional groups that allows attachment to a protein carrier by established measures, so we must modify its structure to introduce such active functional groups. In this experiment, gallium iodide UV lamp was used and acetone as the solvent. The hydrogen-abstraction was followed by radical couplingreaction, cyclic addition reaction between CsA and 4-benzoylbenzoic(BBa) was completed, and CsA-BBa conjugate was obtained in good yield. Activity carboxyl was introduced into CsA molecular. Consequently we coupled CsA-BBa conjugate to poly-L-lysine (PL) by using chemical reaction between amido and carboxyl, and finally acquired needed immunogen. The conformation of the coupling reaction product was confirmed by UV spectra. The coupling efficiency and coverage rate were fairly high and in the optimal recommended range. In this experiment, the coupling product of CsA and PL was even identified by using protein staining and ELISA.Complete immunogen was successfully synthesized. Which not only made the preparation of anti-CsA McAb possibly put into practice, but also provided reference for the immunogen synthesizing of other small molecular substance.