Enthalpy array analysis of enzymatic and binding reactions |
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Authors: | Recht Michael I De Bruyker Dirk Bell Alan G Wolkin Michal V Peeters Eric Anderson Greg B Kolatkar Anand R Bern Marshall W Kuhn Peter Bruce Richard H Torres Frank E |
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Affiliation: | a Scripps-PARC Institute for Advanced Biomedical Sciences, Palo Alto Research Center, 3333 Coyote Hill Rd., Palo Alto, CA 94304, USA b The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA |
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Abstract: | Enthalpy arrays enable label-free, solution-based calorimetric detection of molecular interactions in a 96-detector array format. The combination of the small size of the detectors and the ability to perform measurements in parallel results in a significant reduction of sample volume and measurement time compared with conventional calorimetry. We have made significant improvements in the technology by reducing the temperature noise of the detectors and improving the fabrication materials and methods. In combination with an automated measurement system, the advances in device performance and data analysis have allowed us to develop basic enzyme assays for substrate specificity and inhibitor activity. We have also performed a full titration of 18-crown-6 with barium chloride. These results point to future applications for enthalpy array technology, including fragment-based screening, secondary assays, and thermodynamic characterization of leads in drug discovery. |
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Keywords: | Nanocalorimetry Enzyme assay Binding assay Label-free assay |
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