Recently, German researchers upgraded and optimized mechanization microplate platform of Thermo Fisher Scientific Company, successfully using the the platform for research on biocatalysis, and achieving high-throughput screening of protein engineering mutant library.
Mechanization microplate screening platform has existed for a long time. This platform is widely used in micro upgrade for flat culture, monoclonal antibodies screening and protein crystallization, but the system has not be successfully applied in the field of biological catalysts like directed evolution mutant library screening of enzymes.
The platform is showing as below, including core mechanical arm (Fanuc/Thermo Fisher F5), distributor (nanodrop, Thermo Fisher), four ovens, 96 gun head liquid equipment (Bravo, Agilent Inc.), centrifugal machine (Rotanta 460-Robotic, Hettich GmbH), two microplates ultraviolet - visible light and fluorescence point spectrophotometric meters/readers (Varioskan, ThermoScientific and Omega, BMG, respectively), bar code reader (M3, Microscan), microplate rotating platform (Regrip ThermoScientific) and 96 pin microbial colonial pickers (QPix 420, molecular devices).
A successfully de novo design or an active site plan needs to properly understand all the things, especially theory. As is shown in the diagram here of the library screening case with four directed evolution enzyme mutant. The training on cloning, protein expression and optimization of experimental operation can be well extended to other biological catalyst screenings. They chose four reactions to screen (S) - configuration specific transaminase (ATA), halogenated hydrocarbon dehalogenase (HLD), ring ketone mono oxygenase CHMO and pig acyltransferase (pAcyl), that different multiple mutants were successfully sieved.
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