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High throughput screening for rhMIF HPP tautomerase inhibitors. A, keto-enol tautomerization of HPP catalyzed by rhMIF. B, spectrophotometric detection of rhMIF-catalyzed HPP tautomerase reaction in a 384-well plate format. C, scatter plot of HTS outcome for one of the two replicates of 230,000 compounds. Compounds giving >50% inhibition above the line were classified as hits. D, distribution of the 2,430 hits, categorized into five groups with respect to percent inhibition.
