Immunoanalytical Methods for Screening Vindoline from Catharanthus roseus Cell Cultures

Abstract

An enzyme-linked immunosorbent assay (ELISA) and a radioimmunoassay (RIA) have been developed using antiserum raised against vinblastine-bovine serum albumin. The assays are most sensitive to vindoline, which corresponds to that half of the original dimeric hapten most distant from the coupling site. The measuring range for vindoline in the ELISA is from 10 to 1000 fmol (5-500 pg), while the RIA is about one order of magnitude less sensitive. If the behaviour of the original hapten, vinblastine, is evaluated as cross-reactivity against vindoline in the assays, an average value of 50% is obtained. Cross-reactivities of vincristine and catharanthine were about 1.5% in the ELISA, but in the RIA the cross-reactivity of vincristine was increased to 7% as calculated against vindoline. Application of the assays to the screening of vindoline levels in Catharanthus roseus cultivars and cell cultures is described.