Khoronenkova SV, Tishkov VI
High-throughput screening assay for d-amino acid oxidase. [JOURNAL ARTICLE]
Anal Biochem 2007 Dec 25.
A membrane-based high-throughput screening (HTS) assay for active d-amino acid oxidase (DAAO) in liquid samples as well as in intact Escherichia coli cells has been developed and optimized. The detection limit of the assay was less than 1ng per sample. The method proposed can be used for quantitative DAAO determination in the range of 0.13 to 3.60ng enzyme per probe. The protocol was successfully tested to screen a library of E. coli clones containing mutant DAAOs active toward target substrates.
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