High-throughput fluorescence flow-injection topoisomerase II inhibition assay

A high-throughput fluorescence flow-injection assay is described, suitable for determining the catalytic inhibition of DNA topoisomerase II. The metho d, which separates high molecular mass trypanosome kinetoplast DNA from its decatenated product by centrifugation, should be useful for the rapid and accurate screening of potential anticancer topoisomerase II inhibitors and the determination of their inhibition constants. Advantages of the flow-inj ection method over agarose gel electrophoresis and radioactive centrifugati on assays are that it is faster, more sensitive, highly linear in its respo nse to product formation, and does not require the production of radioactiv e trypanosome kinetoplast DNA substrate. (C) 2001 Elsevier Science BY All r ights reserved.