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About the Authors:
Ehud Goldin
* E-mail: [email protected]
Affiliation: Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Wei Zheng
Affiliation: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Omid Motabar
Affiliations Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America, NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Noel Southall
Affiliation: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Jae Hyuk Choi
Affiliations Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America, NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Juan Marugan
Affiliation: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Christopher P. Austin
Affiliation: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Ellen Sidransky
Affiliation: Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, United States of America
Introduction
High throughput screening (HTS) is widely used for the identification of small molecule leads that can be developed into pharmacological agents. Assay miniaturization in a 1536 well format has made it possible to screen large numbers of compounds at multiple concentrations in primary screens [1]. However, the optimal conditions for implementing this strategy must be tailored individually for each drug target before implementing HTS.
A number of HTS assays have been performed to identify potential lead compounds for several of the lysosomal storage disorders (LSDs) [2], [3], [4], [5]. Almost all of these screens utilized purified recombinant enzyme as the enzyme source, mainly due to the high specificity of the recombinant enzyme, and the availability of large amounts of the enzyme, since several lysosomal enzyme preparations are currently available for enzyme replacement therapy (ERT). In addition, most lysosomal enzymes are hydrolases, which can be formatted into similar fluorogenic enzyme assays. These conditions enable comparisons between the...