title: Comparative analysis of 10 small molecules binding to carbonic anhydrase II by different investigators using Biacore technology.
abstract: In this benchmark study, 26 investigators were asked to characterize the kinetics and affinities of 10 sulfonamide inhibitors binding to the enzyme carbonic anhydrase II using Biacore optical biosensors. A majority of the participants collected data that could be fit to a 1:1 interaction model, but a subset of the data sets obtained from some instruments were of poor quality. The experimental errors in the k(a), k(d), and K(D) parameters determined for each of the compounds averaged 34, 24, and 37%, respectively. As expected, the greatest variation in the reported constants was observed for compounds with exceptionally weak affinity and/or fast association rates. The binding constants determined using the biosensor correlated well with solution-based titration calorimetry measurements. The results of this study provide insight into the challenges, as well as the level of experimental variation, that one would expect to observe when using Biacore technology for small molecule analyses.
authors.affiliation: Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA.
authors.forename: Giuseppe A
authors.initials: GA
authors.lastname: Papalia
authors.forename: Stephanie
authors.initials: S
authors.lastname: Leavitt
authors.forename: Maggie A
authors.initials: MA
authors.lastname: Bynum
authors.forename: Phinikoula S
authors.initials: PS
authors.lastname: Katsamba
authors.forename: Rosemarie
authors.initials: R
authors.lastname: Wilton
authors.forename: Huawei
authors.initials: H
authors.lastname: Qiu
authors.forename: Mieke
authors.initials: M
authors.lastname: Steukers
authors.forename: Siming
authors.initials: S
authors.lastname: Wang
authors.forename: Lakshman
authors.initials: L
authors.lastname: Bindu
authors.forename: Sanjay
authors.initials: S
authors.lastname: Phogat
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Giannetti
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Ryan
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Pudlak
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Matusiewicz
authors.forename: Klaus M
authors.initials: KM
authors.lastname: Michelson
authors.forename: Agnes
authors.initials: A
authors.lastname: Nowakowski
authors.forename: Anh
authors.initials: A
authors.lastname: Pham-Baginski
authors.forename: Jonathan
authors.initials: J
authors.lastname: Brooks
authors.forename: Bryan C
authors.initials: BC
authors.lastname: Tieman
authors.forename: Barry D
authors.initials: BD
authors.lastname: Bruce
authors.forename: Michael
authors.initials: M
authors.lastname: Vaughn
authors.forename: Michael
authors.initials: M
authors.lastname: Baksh
authors.forename: Yun Hee
authors.initials: YH
authors.lastname: Cho
authors.forename: Mieke De
authors.initials: MD
authors.lastname: Wit
authors.forename: Alexandra
authors.initials: A
authors.lastname: Smets
authors.forename: Johan
authors.initials: J
authors.lastname: Vandersmissen
authors.forename: Lieve
authors.initials: L
authors.lastname: Michiels
authors.forename: David G
authors.initials: DG
authors.lastname: Myszka
mesh_terms: D015374:Biosensing Techniques; D002151:Calorimetry; D024402:Carbonic Anhydrase II / Q000737:chemistry* / Q000378:metabolism*; D002257:Carbonic Anhydrase Inhibitors / Q000145:classification / Q000378:metabolism*; D015588:Observer Variation; D011485:Protein Binding; D012108:Research Personnel; D013449:Sulfonamides / Q000037:antagonists & inhibitors* / Q000145:classification; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000592:standards
chemical_list: D002257:Carbonic Anhydrase Inhibitors; D013449:Sulfonamides; D024402:Carbonic Anhydrase II