Comparative analysis of 10 small molecules binding to carbonic anhydrase II by different investigators using Biacore technology

• Published in: Analytical biochemistry 2006-12, Vol.359 (1), p.94-105
• Main Authors: Papalia, Giuseppe A., Leavitt, Stephanie, Bynum, Maggie A., Katsamba, Phinikoula S., Wilton, Rosemarie, Qiu, Huawei, Steukers, Mieke, Wang, Siming, Bindu, Lakshman, Phogat, Sanjay, Giannetti, Anthony M., Ryan, Thomas E., Pudlak, Victoria A., Matusiewicz, Katarzyna, Michelson, Klaus M., Nowakowski, Agnes, Pham-Baginski, Anh, Brooks, Jonathan, Tieman, Bryan C., Bruce, Barry D., Vaughn, Michael, Baksh, Michael, Cho, Yun Hee, Wit, Mieke De, Smets, Alexandra, Vandersmissen, Johan, Michiels, Lieve, Myszka, David G.
• Format: Article
• Language: English
• Subjects: Biacore; Biosensing Techniques; Calorimetry; Carbonic Anhydrase II - chemistry; Carbonic Anhydrase II - metabolism; Carbonic Anhydrase Inhibitors - classification; Carbonic Anhydrase Inhibitors - metabolism; Kinetics; Observer Variation; Protein Binding; Protein–protein interaction; Research Personnel; SPR; Sulfonamides - antagonists & inhibitors; Sulfonamides - classification; Surface plasmon resonance; Surface Plasmon Resonance - instrumentation; Surface Plasmon Resonance - standards
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2006.08.021

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.