Facile method for determination of deoxycytidine kinase activity in biological milieus

Wei-Hua Hao a,c, Li-Chieh Yang a, Jong-Jing Wanga, Chang-Shan Hsu a,
Li-Chien Chang b, Kuang-Yang Hsu c,*
a InnoPharmax Inc., Taipei, Taiwan, ROC
b School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, ROC
c School of Pharmacy, Taipei Medical University, Taipei, Taiwan, ROC

A new analytical method for determining deoxycytidine kinase (dCK) activity in biological milieus using luminescence is reported here. This method, based on utilizing adenosine triphosphate (ATP) as the sole phosphate donor in the kinase reaction and monitoring ATP consumption via a luciferase-based chemiluminescence reaction, is capable of detecting dCK activity without the use of specific substrates or radioisotope techniques. Comparing with the reverse-phase high-performance liquid chromatography method, this new method is suggested to be efficient and sensitive. Further, application of the proposed method for profiling dCK activity in cultured cancer cells revealed that a cervix cell line exhibited the highest dCK activity to gemcitabine metabolism.

Keywords: Chemiluminescent assay, Deoxycytidine kinase, Gemcitabine