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DNA repair enzyme analysis on EWOD fluidic microprocessor

D. Jary, A. Chollat-Namy, Y. Fouillet, J. Boutet, C. Chabrol, G. Castellan, D. Gasparutto and C. Peponnet
CEA, FR

Keywords:
microfluidic, EWOD, DNA repair, microsystem

Abstract:
DNA repair enzyme analysis on EWOD fluidic microprocessor
 
An original analytical assay, aimed at monitoring DNA lesions repair activities has been combined to a dedicated microfluidic EWOD (electrowetting on dielectric) microprocessor.
The quantification of enzymatic repair activities is based on using DNA oligonucleotides, called molecular beacons, containing specific DNA lesion. They are labelled with two fluorophores generating a fluorescent signal if enzymes have performed the DNA base excision. This method is well adapted to screen complex cellular extracts for their ability to repair DNA lesions.
We have designed an EWOD microprocessor comprising a set of elementary fluidic functions for loading, dispensing and mixing reagents.
One DNA repair enzyme was tested with a molecular beacon in a 128nl droplet. A kinetic curve was obtained showing that the microprocessor and the associated fluorescent detection system is perfectly adapted to real time monitoring of such enzymatic activity.
The application field is large: medical diagnostic of illnesses associated with DNA repair deficiencies, identification of cancer predisposition, prediction of individual response to chimio or radio therapy and high throughput screening of repair inhibitors. Indeed, EWOD based actuated system is very versatile and is particularly suited to parallel enzymatic activity tests for the drug discovery industry on restricted amount of cellular extracts.

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