Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators.
Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators.
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DOI：
10.1021/ac300478f
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发表时间：
2012-07-03
影响因子：
7.4
通讯作者：
Sligar, Stephen G.
中科院分区：
文献类型：
--
作者：
Marty, Michael T.;Sloan, Courtney D. Kuhnline;Bailey, Ryan C.;Sligar, Stephen G.
探测生物传感器表面大分子结合动力学的常规方法采用逐步滴定分析物浓度，并测量在每个浓度水平下与固定化配体的缔合和解离。以前已经表明，动力学速率可以通过监测结合在一个单一的步骤，随着分析物浓度随时间的线性梯度增加。我们在这里报告的应用程序的非线性分析物浓度梯度确定动力学速率和平衡结合亲和力在一个单一的实验。提出了一种适用于微流控系统的非线性梯度标记器。模拟验证，准确的动力学速率可以提取广泛的关联和解离速率，梯度斜率和曲率，并与模型的质量传输。非线性分析物梯度法证明了与硅光子微谐振器平台，以测量前列腺特异性抗原-抗体结合动力学。
Conventional methods to probe the binding kinetics of macromolecules at biosensor surfaces employ a stepwise titration of analyte concentrations and measure the association and dissociation to the immobilized ligand at each concentration level. It has previously been shown that kinetic rates can be measured in a single step by monitoring binding as the analyte concentration increases over time in a linear gradient. We report here the application of nonlinear analyte concentration gradients for determining kinetic rates and equilibrium binding affinities in a single experiment. A versatile nonlinear gradient maker is presented, which is easily applied to microfluidic systems. Simulations validate that accurate kinetic rates can be extracted for a wide range of association and dissociation rates, gradient slopes and curvatures, and with models for mass transport. The nonlinear analyte gradient method is demonstrated with a silicon photonic microring resonator platform to measure prostate specific antigen-antibody binding kinetics.
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影响因子：
6.1
作者：
Choi, Charles J.;Cunningham, Brian T.
通讯作者：
Cunningham, Brian T.
影响因子：
2.9
作者：
Karlsson, R;Katsamba, PS;Myszka, DG
通讯作者：
Myszka, DG
影响因子：
2.9
作者：
Quinn, John G.
通讯作者：
Quinn, John G.
影响因子：
3.4
作者：
Myszka, DG;He, X;Goldstein, B
通讯作者：
Goldstein, B
DOI：
10.1006/meth.1994.1013
发表时间：
1994-01-01
期刊：
Methods (Orlando)
影响因子：
--
作者：
Karlsson, Robert;Roos, Hakan;Persson, Bjorn
通讯作者：
Persson, Bjorn