Development of an automated assay for accelerated in vitro detection of DNA adduct-inducing and crosslinking agents
Jelena Medan, Brad E Sleebs, Kurt Lackovic, Keith G Watson, Benny J Evison, Don R Phillips, Suzanne M Cutts
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS | PERGAMON-ELSEVIER SCIENCE LTD | Published : 2021
Current techniques for the identification of DNA adduct-inducing and DNA interstrand crosslinking agents include electrophoretic crosslinking assays, electrophoretic gel shift assays, DNA and RNA stop assays, mass spectrometry-based methods and 32P-post-labelling. While these assays provide considerable insight into the site and stability of the interaction, they are relatively expensive, time-consuming and sometimes rely on the use of radioactively-labelled components, and thus are ill-suited to screening large numbers of compounds. A novel medium throughput assay was developed to overcome these limitations and was based on the attachment of a biotin-tagged double stranded (ds) oligonucleot..View full abstract
Awarded by National Health and Medical Research Council, Australia
Awarded by CASS Foundation, Australia
We thank Patrizia Novello for technical assistance with automation. This work was supported by grants from the National Health and Medical Research Council, Australia [grant number 487333] and the CASS Foundation, Australia [grant numbers SM/07/1547 and SM/08/1971]. B.E.S. is an Ellen Corin Fellow.