Kinetic analysis and fragment screening with Fujifilm AP-3000.
We evaluated the performance of Fujifilm's new AP-3000 surface plasmon resonance biosensor for kinetic analysis and fragment screening. Using carbonic anhydrase II as a model system, we characterized a set of 10 sulfonamide-based inhibitors that range in molecular mass from 98 to 341Da and approximately 10,000-fold in affinity (0.4mM to 20nM). Although the data collected from the AP-3000 were generally similar to those collected using a Biacore T100, the AP-3000's stop-flow analyte delivery system complicated the shapes of the association- and dissociation-phase binding responses. We illustrate how reasonable estimates of the kinetic rate constants can be extracted from AP-3000 data by limiting data analysis to only the regions of the responses collected during flow conditions. We also provide an example of the results obtained for a fragment-screening study with the AP-3000, which is the ideal application of this technology.
Rich
Rebecca L
RL
Center for Biomolecular Interaction Analysis, University of Utah School of Medicine, Salt Lake City, 84132, USA.
Myszka
David G
DG
D024402:Carbonic Anhydrase II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002257:Carbonic Anhydrase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods; D007700:Kinetics; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology*; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods
D002257:Carbonic Anhydrase Inhibitors; D013449:Sulfonamides; D024402:Carbonic Anhydrase II