Facile method for determination of deoxycytidine kinase activity in biological milieus

Journal of Food and Drug Analysis

Wei-Hua Hao¹  Jong-Jing Wang¹  Li-Chieh Yang¹  Chang-Shan Hsu¹  Li-Chien Chang²  Kuang-Yang Hsu³ 

[1] InnoPharmax Inc., Taipei, Taiwan, ROC;School of Pharmacy, National Defense Medical Center, Taipei, Taiwan, ROC;School of Pharmacy, Taipei Medical University, Taipei, Taiwan, ROC;

关键词: Chemiluminescent assay;    Deoxycytidine kinase;    Gemcitabine;   

DOI  :  10.1016/j.jfda.2013.09.008

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown