DNA repair enzyme analysis on EWOD fluidic microprocessor

摘要

An original homogeneous phase assay, aimed at monitoring DNA lesions repair activities, has been combined to a microfluidic EWOD (electrowetting on dielectric) microprocessor. Such miniaturized devices are particularly advantageous when parallel analyses must be performed with a very limited sample volume. This is the case for the diagnostic of diseases or drug discovery associated to the cellular DNA repair activity.We have compared the activity of repair enzyme (Fpg) in 120 nl droplets on chip and in 6μl samples in tube with real time fluorescence detection. No loss of enzyme activity was measured for the limited on chip volumes tested. Sensitivity of the test was equivalent in tube and on chip, with 5nM of enzyme detected. Finally, we have shown that proteic cellular extract are reproducibly dispensed and mixed on chip by EWOD actuation.These validations are very promising for DNA repair test performed on chip with cellular extract for diagnostic applications.