Polymerase assays for lead discovery: An overall review of methodologies and approaches.

Abstract Polymerases represent an attractive molecular target for antibacterial drug development, antiviral intervention and cancer therapy. Over the past decade, academic groups and scientists from pharmaceutical industry have developed a large plethora of different functional assays to monitor the enzymatic reaction catalyzed by polymerases. These assays were used to enable high-throughput screening (HTS) for lead discovery purposes, as well as hit-to-lead (H2L) drug profiling activities. In both cases the choice of the assay technology is critical and to the best of our knowledge, there is no review available to help scientists to choose the most suitable assay. This review summarizes the most common functional assays developed to monitor the enzymatic activity of polymerases and discusses the advantages and disadvantages of each assay. Assays are presented and evaluated in term of cost, ease of use, high-throughput screening compatibility and liability towards delivering false positives and false negatives. Highlights • Overview on functional assays to monitor the enzymatic reaction of polymerases. • Application of assays in high-throughput screening (HTS) for lead discovery. • Recommendation for choosing the appropriate assay technology. • Evaluation of assays in term of cost, ease of use and HTS compatibility. • Application of assays for antibacterial, antiviral anticancer drug development. [ABSTRACT FROM AUTHOR]