title: Thermodynamic benchmark study using Biacore technology.
abstract: A total of 22 individuals participated in this benchmark study to characterize the thermodynamics of small-molecule inhibitor-enzyme interactions using Biacore instruments. Participants were provided with reagents (the enzyme carbonic anhydrase II, which was immobilized onto the sensor surface, and four sulfonamide-based inhibitors) and were instructed to collect response data from 6 to 36 degrees C. van't Hoff enthalpies and entropies were calculated from the temperature dependence of the binding constants. The equilibrium dissociation and thermodynamic constants determined from the Biacore analysis matched the values determined using isothermal titration calorimetry. These results demonstrate that immobilization of the enzyme onto the sensor surface did not alter the thermodynamics of these interactions. This benchmark study also provides insights into the opportunities and challenges in carrying out thermodynamic studies using optical biosensors.
authors.affiliation: Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA.
authors.forename: Iva
authors.initials: I
authors.lastname: Navratilova
authors.forename: Giuseppe A
authors.initials: GA
authors.lastname: Papalia
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Rich
authors.forename: Daniel
authors.initials: D
authors.lastname: Bedinger
authors.forename: Susan
authors.initials: S
authors.lastname: Brophy
authors.forename: Brad
authors.initials: B
authors.lastname: Condon
authors.forename: Ta
authors.initials: T
authors.lastname: Deng
authors.forename: Anne W
authors.initials: AW
authors.lastname: Emerick
authors.forename: Hann-Wen
authors.initials: HW
authors.lastname: Guan
authors.forename: Tanya
authors.initials: T
authors.lastname: Hayden
authors.forename: Thomas
authors.initials: T
authors.lastname: Heutmekers
authors.forename: Bart
authors.initials: B
authors.lastname: Hoorelbeke
authors.forename: Mark C
authors.initials: MC
authors.lastname: McCroskey
authors.forename: Mary M
authors.initials: MM
authors.lastname: Murphy
authors.forename: Terry
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Fabio
authors.initials: F
authors.lastname: Parmeggiani
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Qin
authors.forename: Sabina
authors.initials: S
authors.lastname: Rebe
authors.forename: Nenad
authors.initials: N
authors.lastname: Tomasevic
authors.forename: Tiffany
authors.initials: T
authors.lastname: Tsang
authors.forename: M Brett
authors.initials: MB
authors.lastname: Waddell
authors.forename: Fred Feiyu
authors.initials: FF
authors.lastname: Zhang
authors.forename: Stephanie
authors.initials: S
authors.lastname: Leavitt
authors.forename: David G
authors.initials: DG
authors.lastname: Myszka
mesh_terms: D019985:Benchmarking; D035843:Biomedical Research; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000592:standards; D002151:Calorimetry / Q000295:instrumentation* / Q000592:standards*; D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000378:metabolism; D002257:Carbonic Anhydrase Inhibitors / Q000145:classification* / Q000378:metabolism*; D015588:Observer Variation; D011485:Protein Binding; D013449:Sulfonamides / Q000037:antagonists & inhibitors* / Q000145:classification; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000592:standards; D013816:Thermodynamics
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D013449:Sulfonamides; D024402:Carbonic Anhydrase II