Quantitative measurements and modeling of kinetics in nucleic acid monolayer films using SPR spectroscopy
Quantitative measurements and modeling of kinetics in nucleic acid monolayer films using SPR spectroscopy
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DOI：
10.1021/ja9930824
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发表时间：
2000-04-05
影响因子：
15
通讯作者：
Peterson, AW
中科院分区：
文献类型：
--
作者：
Georgiadis, R;Peterlinz, KP;Peterson, AW
我们报告了一个定量研究动力学的形成双组分系链ssDNA单层膜使用原位双色表面等离子体共振（SPR）光谱。DNA与金的连接是通过在5'端与巯基（HS-C-6-ssDNA）连接的功能化而实现的。详细的数据分析是通过定量比较从SPRS获得的DNA覆盖率与时间动力学数据，以及在界面上同时吸附、解吸和扩散的微分方程的数值解li，r进行的。HS-C-6-ssDNA在裸金上的吸附动力学以及随后巯基己醇处理过程中HS-C-6-ssDNA从表面损失的动力学可以用一个简单的物理模型和自一致的参数来理解。将HS-C-6-ssDNA在裸金上的吸附动力学与表面附着的巯基化ssDNA与自由溶液中完全互补的ssDNA的杂交动力学进行了比较，发现它们遵循非常相似的动力学途径。相反，ssDNA的吸附遵循复杂的动力学，不能用单一的动力学步骤来建模。也就是说，在25-mer的ssDNA上存在巯基功能会导致吸附行为，这种吸附行为在动力学上明显不同于简单的ssDNA在金上的吸附。
We report a quantitative study of the kinetics of formation for a two-component tethered ssDNA monolayer film using in situ two-color surface plasmon resonance (SPR) spectroscopy. The attachment of the DNA to gold is facilitated by functionalization at the 5' end with a thiol group connected by a hexamethylene linker (HS-C-6-ssDNA). Detailed data analysis is performed by quantitative comparison of the DNA coverage versus time kinetic data obtained from SPRS with numerical solutions li,r the differential equations for simultaneous adsorption, desorption, and diffusion at the interface. The kinetics of adsorption of HS-C-6-ssDNA onto bare gold as well as the kinetics of loss of HS-C-6-ssDNA from the: surface during subsequent treatment with mercaptohexanol can be understood in terms of a simple physical model and self-consistent parameters. The kinetics of HS-C-6-ssDNA adsorption on bare gold are compared to the kinetics of hybridization of surface-attached thiolated ssDNA with the fully complementary ssDNA in free solution and found to follow remarkably similar kinetic pathways. In contrast, the adsorption of ssDNA follows complex kinetics that cannot be modeled with a single kinetic step. That is, the presence of a thiol functionality on a 25-mer ssDNA gives rise to adsorption behavior that is clearly kinetically distinct from simple ssDNA adsorption on gold.