Extending the range of rate constants available from BIACORE: Interpreting mass transport-influenced binding data
Extending the range of rate constants available from BIACORE: Interpreting mass transport-influenced binding data
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DOI：
10.1016/s0006-3495(98)77549-6
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发表时间：
1998-08-01
影响因子：
3.4
通讯作者：
Goldstein, B
中科院分区：
文献类型：
--
作者：
Myszka, DG;He, X;Goldstein, B
基于表面的结合测定通常受分析物到传感器表面的运输的影响。使用模拟数据集，我们测试了一个简单的两室模型，看看它的描述运输和绑定是足够的准确分析BIACORE数据。首先，我们提出了一个计算机模型，可以产生现实的BIACORE数据。该模型计算分析物在流动池内的层流，其垂直和平行于传感器表面的扩散，以及分析物和固定化反应物之间的可逆化学反应。我们使用这个计算机模型来生成各种条件下的绑定数据。这些数据集与二室模型的分析表明，良好的估计的内在反应速率常数恢复，即使当质量传输影响的结合反应。我们还讨论了条件下，可以使用二室模型来确定的分析物的扩散系数。我们的研究结果表明，该模型可以显着扩展的范围内的关联速率常数，可以准确地确定从BIACORE。
Surface-based binding assays are often influenced by the transport of analyte to the sensor surface. Using simulated data sets, we test a simple two-compartment model to see if its description of transport and binding is sufficient to accurately analyze BIACORE data. First we present a computer model that can generate realistic BIACORE data. This model calculates the laminar flow of analyte within the flow cell, its diffusion both perpendicular and parallel to the sensor surface, and the reversible chemical reaction between analyte and immobilized reactant. We use this computer model to generate binding data under a variety of conditions. An analysis of these data sets with the two-compartment model demonstrates that good estimates of the intrinsic reaction rate constants are recovered even when mass transport influences the binding reaction. We also discuss the conditions under which the two-compartment model can be used to determine the diffusion coefficient of the analyte. Our results illustrate that this model can significantly extend the range of association rate constants that can be accurately determined from BIACORE.