Roden L D, Myszka D G
Oncological Sciences Department, Huntsman Cancer Institute, University of Utah, Salt Lake City 84112, USA.
Biochem Biophys Res Commun. 1996 Aug 23;225(3):1073-7. doi: 10.1006/bbrc.1996.1297.
We demonstrate that the interaction between myoglobin and an immobilized anti-myoglobin antibody measured on BIAcore 2000 can be described by a simple bimolecular reaction mechanism. We improved the quality of the sensor data by correcting for refractive index changes and nonspecific binding using a blank sensor surface. Applying nonlinear least squares analysis, we simultaneously fitted the association and dissociation phase data generated for a range of myoglobin concentrations injected across the antibody surface. The ability to globally fit these data to a simple binding model indicates that effects related to the sensor surface, like mass transport and the dextran matrix, did not complicate the observed binding responses. These results illustrate the potential of BIAcore to monitor macromolecular interactions in real-time and the utility of global analysis to resolve the reaction kinetics.
我们证明，在BIAcore 2000上测量的肌红蛋白与固定化抗肌红蛋白抗体之间的相互作用可用简单的双分子反应机制来描述。我们通过使用空白传感器表面校正折射率变化和非特异性结合来提高传感器数据的质量。应用非线性最小二乘法分析，我们同时拟合了在抗体表面注入一系列肌红蛋白浓度时产生的结合和解离阶段数据。将这些数据整体拟合到一个简单结合模型的能力表明，与传感器表面相关的效应，如质量传输和葡聚糖基质，并未使观察到的结合反应复杂化。这些结果说明了BIAcore实时监测大分子相互作用的潜力以及整体分析在解析反应动力学方面的实用性。
