Nathalie Maubon, Christelle Valaire, Séverine Michelin and Philippe Masson. (Abbott Group, Laboratoires Fournier, Research Center, Global System Biology Department, HCS unit, 50 rue de Dijon, 21121 DAIX, France)
Development of a fully automated transactivation assay using the CompacT selecT.
Laboratoires Fournier has extensive expertise in nuclear receptors analysis. Several screening runs have been performed, like PPAR or LXR, using transactivation assay on Cos-7 cells. In previous screenings, cell culture, transfection and plating were performed using a semi-automated mode whereas cell treatments with drugs and read-out were fully automated. To improve the throughput, full automation was envisaged.
In this context, Laboratoires Fournier has acquired the CompacT SelecT, an advanced cell culture automated system from The Automation Partnership. This system allows cell production in single or triple flasks up to 90 flasks and plating cells automatically in 384-well plates up to 210 plates.
3 different conditions for study of agonists dose response effect on LXR and Z and Z' analysis were tested during the assay development phase. These were as follows:
- manual cell culture, transfection and plating,
- cell culture on the automated system and manual transfection and plating,
- cell culture, transfection and plating on the automated system.
The EC50 curves were satisfactorily reproducible in the 3 different situations. Furthermore, Z and Z' were not affected by full automation.
In conclusion, a high-throughput fully automated transactivation assay has been developed, allowing increased throughput, reduction in turnaround and better productivity.
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