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Miniaturization of an Enzyme Assay (β-Galactosidase) in the 384- and 1536-Well Plate Format

1999; SAGE Publishing; Volume: 4; Issue: 6 Linguagem: Inglês

10.1016/s1535-5535(04)00042-5

1540-2452

Michael G. Berg, Katrin Undisz, Ralf Thiericke, Thomas C. Moore, Clemens Posten,

Advanced Biosensing Techniques and Applications

Miniaturization is one way to realize today's demands in the drug discovery process by moving from the standard 96-well plate to higher density microplate formats. In this article we describe the adaptation of a fluorescence-based enzyme assay to the challenges of the 384- and 1536-well plate format. The liquid-handling was realized by the automated micropipettor CyBi-Well™ 96/384/1536* (CyBio AG — formerly JENOPTIK Bioinstruments Gmbh — Jena, Germany). On the basis of optimized liquid-handling parameters pipetting routines were established to perform an enzyme assay (β-galactosidase) in the microplate formats of higher density. Finally, the experimental results were compared to those obtained in the well-established 96-well format. In the enzyme assay, the bioconversion of the substrate Fluorescein-di-(β-D-galactopyranoside) (FDG), occurred as a linear function of the β-galactosidase concentration comparably in all three assay formats. We conclude that miniaturization using the higher density 384- and 1536-well plate formats is advantageous as the next evolutionary step in HTS, especially using enzyme assays. A careful individual adaptation procedure for each microplate format and assay at the basis of the optimized liquid-handling parameters is essential. CyBi-Well™ 96/384/1536 proves to be a powerful tool for a careful adaptation of the liquid-handling procedures of biological assays especially also in the 384- and 1536-well formats.