New taxol assay method using tubulin-assembly

Abstract

A novel taxol determination method which; involves the tubulin-assembly stimulation is described. The tubulin-assembly was monitored by turbidity change at 350 nm. In a limited range of taxol concentration (0 to 24 mu M), taxol stimulated tubulin-assembly linearly. And this linear relation was observed from 20 min to 30 min after the reaction started. Bioactive derivatives of taxol such as cephalomanin and 7-epi-10-deacetyltaxol also stimulated the tubulin assembly. However, baccatin III, which was known as less active taxol derivative did not stimulate tubulin assembly. This result showed that the stimulation of tubulin assembly has a relationship with the antimiotic activity. This assay method have several advantages. 1) Time required for the measurement is relatively short. 2) Multiple samples can be measured simultaneously. 3) It can remove interference of less active taxane compounds more selectively than immuno-assay. Consequently, this method can be used to determine taxol concentration in biological samples. Especially, this method can be used for large scale selection of cell line and primary screening of new antimiotic compounds.