Wu Jun, Wang Hongliang, Zhu Anna, Long Feng
School of Environment and Natural Resources, Renmin University of China, Beijing 10085, China.
ACS Omega. 2018 May 31;3(5):5605-5614. doi: 10.1021/acsomega.7b02063. Epub 2018 May 25.
Thorough understandings on the real-time kinetics involved in DNA adsorption on a solid surface is essential in various fields, such as in DNA hybridization studies, DNA extraction and purification, DNA-based biosensing, and gene-based medicine discovery. Herein, the real-time properties of single-stranded DNA (ssDNA) adsorption onto functional silica surfaces under various conditions were investigated using an evanescent wave optical biosensing platform. Results demonstrated that the driving force and adsorption mechanism of DNA were closely related to the kind of functional groups on the silica surfaces. The main driving forces of DNA adsorption onto hydroxyl- and protein-modified solid surfaces were the hydrophobic interaction, hydrogen bonding, and the interaction between DNA phosphate and functional groups on the silica surface, which strengthened with increased ionic strength. However, the electrostatic attraction between the negative charge of DNA and positive charge of the amino silica surface was likely the most important factor influencing DNA adsorption onto the amino surface. This influence can be reduced by increasing the ionic strength. Although low-ionic-strength Mg provided a greater adsorption efficiency than high-ionic-strength Na, the balance of ssDNA adsorption onto hydroxyl- and ovalbumin (OVA)-modified silica surfaces was achieved faster in the presence of Na than in the presence of Mg. DNA adsorption was also influenced significantly by pH, and the hydroxyl- and OVA-modified surfaces exhibited the strongest adsorption at pH 3.0, whereas DNA adsorption onto the amino surface increased with increased pH. DNA adsorption onto various functional surfaces could be perfectly fitted by second-order Langmuir models, indicating that the process was a single-molecular-layer adsorption.
深入了解DNA在固体表面吸附的实时动力学在多个领域至关重要,例如DNA杂交研究、DNA提取与纯化、基于DNA的生物传感以及基于基因的药物发现。在此,利用倏逝波光学生物传感平台研究了单链DNA(ssDNA)在各种条件下吸附到功能化二氧化硅表面的实时特性。结果表明,DNA的驱动力和吸附机制与二氧化硅表面官能团的种类密切相关。DNA吸附到羟基和蛋白质修饰的固体表面的主要驱动力是疏水相互作用、氢键以及DNA磷酸基团与二氧化硅表面官能团之间的相互作用,这些作用会随着离子强度的增加而增强。然而,DNA的负电荷与氨基二氧化硅表面的正电荷之间的静电吸引可能是影响DNA吸附到氨基表面的最重要因素。通过增加离子强度可以降低这种影响。尽管低离子强度的Mg比高离子强度的Na具有更高的吸附效率,但在存在Na的情况下,ssDNA吸附到羟基和卵清蛋白(OVA)修饰的二氧化硅表面的平衡比在存在Mg的情况下更快实现。DNA吸附也受到pH的显著影响,羟基和OVA修饰的表面在pH 3.0时表现出最强的吸附,而DNA在氨基表面的吸附随着pH的升高而增加。DNA在各种功能表面上的吸附可以用二阶朗缪尔模型完美拟合,表明该过程是单分子层吸附。