DEVELOPMENT OF ENZYME-IMMUNOASSAY OF 2'-DEOXYCYTIDINE

In order to study 2'-deoxycytidine (2'-dCyd) as a possible prognostic marker in cancer chemotherapy, an enzyme immunoassay (EIA) was develop ed. 2'-dCyd as a hapten was succinylated and two O-monosuccinyl-2'-dCy d's were purified by high performance liquid chromatography and identi fied by mass spectrometry and H-1-NMR. Two antigens were prepared by c onjugating two O-succinyl derivatives with keyhole limpet hemocyanin ( KLH) as a carrier. Both antigen produced specific antibodies to 2'-dCy d in BALB/c mice. The spleen cells of one mouse immunized with 5'-O-su ccinyl-2'-dCyd-KLH were hybridized with myeloma cells. One monoclone s elected produced a specific antibody. A convenient EIA was attained by using the monoclonal antibody. (C) 1997 Elsevier Science B.V.