Extracting kinetic rate constants from surface plasmon resonance array systems

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• 作者：Rebecca L. Rich ; Michelle J. Cannon ; Jerry Jenkins ; Prabhakar P ; ian ; Shankar Sundaram ; Rachelle Magyar ; Jennifer Brockman ; Jeremy Lambert ; David G. Myszka
• 关键词：Biacore ; Protein– ; protein interaction ; Affinity ; Flexchip
• 刊名：Analytical Biochemistry
• 出版年：2008
• 出版时间：1 February 2008
• 年：2008
• 卷：373
• 期：1
• 页码：112-120
• 全文大小：1695 K

文摘

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.