Synthesis Of The Bis-analogue And The Immunogen Of Cyclosporin A

In this paper the principle of sandwich enzyme immunoassay of cyclosporin A (CsA) using bis-analogue of the CsA has been described. Furthermore, the bis-analogue of CsA and the immunogen of CsA, which are the necessary material to establish the promising analytical technique, have been prepared.The route we designed to prepare the bis-analogue of the CsA was as follows: CsA reacted with 4-benzoylbenzoic acid under ultraviolet radiation to generate the conjugate of CsA-BBa, which has a reactive carboxyl. Then, the bis-analogue of CsA was prepared by the reaction between the conjugate of CsA-BBa and ethylenediamine with the action of 2-bromo-l-ethyl pyridinium tetrafluoroborate (BEP). As a result, we synthesized the bis-analogue of the CsA successfully in laboratory.hi the experiment to synthesize the immunogen of CsA, l-ethyl-3-(3-dimethylamino propyl)-carbodiimide hydrochloride (EDCI) was selected as a coupling reagent; the conjugate of CsA-BBa was linked to poly-L-lysine to generate CsA immunogen. When CsA-BBa-PL was tested by indirect ELISA, the specific affinity reaction occurred between the conjugate of CsA-BBa-PL and the monoclonal antibodies to CsA. It suggested that the methods of antigen synthesis were practicable.In the research to prepare the bis-analogue of CsA and the immunogen of CsA,the mechanism of the photochemistry and coupling reaction had been discussed on the basis of the experimental detail and related knowledge.In order to develop the sandwich enzyme immunoassay of CsA, the bis-analogue of CsA and the immunogen of CsA were prepared. The corresponding method and technology will throw the light on other, especially, the hapten lacking in reactive functional groups.