Extracting kinetic rate constants from surface plasmon resonance array systems.
Surface plasmon resonance imaging systems, such as Flexchip from Biacore, are capable of monitoring hundreds of reaction spots simultaneously within a single flow cell. Interpreting the binding kinetics in a large-format flow cell presents a number of potential challenges, including accounting for mass transport effects and spot-to-spot sample depletion. We employed a combination of computer simulations and experimentation to characterize these effects across the spotted array and established that a simple two-compartment model may be used to accurately extract intrinsic rate constants from the array under mass transport-limited conditions. Using antibody systems, we demonstrate that the spot-to-spot variability in the binding kinetics was <9%. We also illustrate the advantage of globally fitting binding data from multiple spots within an array for a system that is mass transport limited.
Rich
Rebecca L
RL
Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA.
Cannon
Michelle J
MJ
Jenkins
Jerry
J
Pandian
Prabhakar
P
Sundaram
Shankar
S
Magyar
Rachelle
R
Brockman
Jennifer
J
Lambert
Jeremy
J
Myszka
David G
DG
D007700:Kinetics; D008024:Ligands; D008962:Models, Theoretical; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
D008024:Ligands