Enthalpy array analysis of enzymatic and binding reactions.
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.
Recht
Michael I
MI
Scripps-PARC Institute for Advanced Biomedical Sciences, Palo Alto Research Center, 3333 Coyote Hill Rd., Palo Alto, CA 94304, USA. mrecht@parc.com
De Bruyker
Dirk
D
Bell
Alan G
AG
Wolkin
Michal V
MV
Peeters
Eric
E
Anderson
Greg B
GB
Kolatkar
Anand R
AR
Bern
Marshall W
MW
Kuhn
Peter
P
Bruce
Richard H
RH
Torres
Frank E
FE
D001331:Automation / Q000295:instrumentation; D017609:Barium Compounds / Q000378:metabolism; D002151:Calorimetry / Q000295:instrumentation / Q000379:methods*; D002712:Chlorides / Q000378:metabolism; D043844:Crown Ethers / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D004798:Enzymes / Q000378:metabolism*; D006593:Hexokinase / Q000378:metabolism; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D011485:Protein Binding; D013379:Substrate Specificity; D013816:Thermodynamics; D017186:Titrimetry; D014361:Trypsin Inhibitors / Q000378:metabolism / Q000494:pharmacology
D017609:Barium Compounds; D002712:Chlorides; D043844:Crown Ethers; D004791:Enzyme Inhibitors; D004798:Enzymes; D014361:Trypsin Inhibitors; C024986:barium chloride; C015762:18-crown-6; D006593:Hexokinase