Assay high-throughput approach

Fig. 13.8. Purification of 96 different enzyme variants in a high-throughput approach. Cells are grown, harvested and lysed in microplates. His6-tagged variants are isolated by transferring the lysate to Ni-NTA coated microplates from which they are eluted after washing. Specific enzyme activity can be checked by suitable assays.

In a different approach a super-high-throughput ee-assay was developed on the basis of chirally modified capillary array electrophoresis (CAE).90 CAE was used in the Human Genome Project, and commercially available instruments have been developed which comprise a high number of capillaries in parallel, for example the 96-capillary unit MegaBACE consisting of 6 bundles of 16 capillaries.91 The system can address a 96-well microtiter plate. It was adapted to perform ee-determinations of chiral amines, which are potentially accessible by catalytic reductive amination of ketones, transition metal catalyzed Markovnikov addition of ammonia, or enzymatic hydrolysis of acetamides (Scheme 14).90... 

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